{"title":"3D Organoid Culture 1.2","description":"\u003cp\u003e\u003cspan\u003e3D Organoid Culture 1.2\u003c\/span\u003e\u003c\/p\u003e","products":[{"product_id":"mammary-organoids-cytokine-set-human","title":"Mammary organoids Cytokine Set, Human","description":"\u003ch4\u003eProduct Specification\u003c\/h4\u003e\n\u003cdiv class=\"responsive-table product-detail-table details-table\"\u003e\n\u003cbr\u003e\n\u003ctable style=\"width: 100%; height: auto;\"\u003e\n\u003ctbody\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 22%;\"\u003e\u003cstrong\u003eSpecies\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 78%;\"\u003eHuman\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 22%;\"\u003e\u003cstrong\u003eSynonyms\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 78%;\"\u003eMammary organoids Cytokine Set\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 22%;\"\u003e\u003cstrong\u003ePhysical Appearance\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 78%;\"\u003eLyophilized Powder\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 22%;\"\u003e\u003cstrong\u003eStability \u0026amp; Storage\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 78%;\"\u003e\n\u003cp\u003e· 12 months from date of receipt, lyophilized powder stored at -20 to -80℃. \u003cbr\u003e· 3 months, -20 to -80℃ under sterile conditions after reconstitution.\u003cbr\u003e· 1 week, 2 to 8℃ under sterile conditions after reconstitution. \u003cbr\u003e· Please avoid repeated freeze-thaw cycles.\u003c\/p\u003e\n\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003c\/tbody\u003e\n\u003c\/table\u003e\n\u003c\/div\u003e\n\u003ch4\u003eComponents\u003c\/h4\u003e\n\u003cdiv\u003e\n\u003cp\u003eS-10μg\u003cbr\u003eM-50μg\u003cbr\u003eL-100μg\u003c\/p\u003e\n\u003c\/div\u003e","brand":"UA BIOSCIENCE","offers":[{"title":"S","offer_id":41519875063883,"sku":null,"price":1056.0,"currency_code":"USD","in_stock":true},{"title":"M","offer_id":41519875096651,"sku":null,"price":2690.0,"currency_code":"USD","in_stock":true},{"title":"L","offer_id":41519875129419,"sku":null,"price":4544.0,"currency_code":"USD","in_stock":true}],"thumbnail_url":"\/\/cdn.shopify.com\/s\/files\/1\/0590\/8375\/1499\/files\/AntBioImage_b0825f42-8837-46d0-92ae-8afde80f8ac3.png?v=1770715627"},{"product_id":"lung-organoids-cytokine-setmouse-ua090041","title":"Lung organoids Cytokine Set,Mouse","description":"\u003ch4\u003eProduct Specification\u003c\/h4\u003e\u003cdiv class=\"responsive-table product-detail-table details-table\"\u003e\n\u003cbr\u003e\u003ctable style=\"width: 100%; height: auto;\"\u003e\u003ctbody\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 22%;\"\u003e\u003cstrong\u003eEndotoxin\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 78%;\"\u003e\u0026lt;0.1EU\/μg\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 22%;\"\u003e\u003cstrong\u003ePhysical Appearance\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 78%;\"\u003eLyophilized powder\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 22%;\"\u003e\u003cstrong\u003eReconstitution\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 78%;\"\u003e\u003cp style=\"margin-bottom: 0px;\"\u003eReconstitute at 0.1-1 mg\/ml according to the size in ultrapure water after rapid centrifugation.\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 22%;\"\u003e\u003cstrong\u003eStability \u0026amp; Storage\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 78%;\"\u003e\u003cp\u003e·12 months from date of receipt, lyophilized powder stored at -20 to -80℃. \u003cbr\u003e· 3 months, -20 to -80℃ under sterile conditions after reconstitution. \u003cbr\u003e· 1 week, 2 to 8℃ under sterile conditions after reconstitution. \u003cbr\u003e· Please avoid repeated freeze-thaw cycles.\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003c\/tbody\u003e\u003c\/table\u003e\n\u003c\/div\u003e\u003ch4\u003eBackground\u003c\/h4\u003e\u003cdiv\u003e\u003cp\u003eThe mouse lung organoid cell factor kit is based on an optimized formulation derived from research, and contains key factors such as Wnt3a and FGF10 that have been verified in the literature. It is specifically designed for the efficient construction of lung organoids and is suitable for studies on development, diseases, and drug screening. Mouse lung organoids are three-dimensional (3D) micro-lung tissue models cultivated in vitro, composed of mouse lung epithelial stem cells (such as basal cells, type II alveolar cells), and have characteristics including the retention of the cell types of the source tissue (such as alveolar epithelial cells, bronchial cells) and some functions (such as surfactant secretion). They simulate lung development, regeneration, and disease processes, compensating for the limitations of traditional 2D cell models and animal models.\u003cbr\u003e\u003c\/p\u003e\u003c\/div\u003e\u003ch4\u003eComponents\u003c\/h4\u003e\u003cdiv\u003e\u003ctable class=\"table table-bordered\" style=\"width: 646.41px;\"\u003e\u003ctbody\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 138.669px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:11.0000pt\"\u003e\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 192.012px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:10.0000pt\"\u003eReference dosage\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 159.009px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:10.0000pt\"\u003e100ml \u003cfont\u003esystem\u003c\/font\u003e\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 156.005px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:10.0000pt\"\u003e1000ml \u003cfont\u003esystem\u003c\/font\u003e\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 298.218px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:11.0000pt\"\u003e\u003cfont\u003eCytokines1\u003c\/font\u003e\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 298.218px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:10.0000pt\"\u003e50ng\/ml\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 298.218px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:10.0000pt\"\u003e5μg\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 298.218px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:10.0000pt\"\u003e50μg\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 298.218px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:11.0000pt\"\u003e\u003cfont\u003eCytokines2\u003c\/font\u003e\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 298.218px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:10.0000pt\"\u003e500ng\/ml\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 298.218px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:10.0000pt\"\u003e50μg\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 298.218px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:10.0000pt\"\u003e500μg\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 298.218px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:11.0000pt\"\u003e\u003cfont\u003eCytokines3\u003c\/font\u003e\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 298.218px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:10.0000pt\"\u003e100ng\/ml\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 298.218px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:10.0000pt\"\u003e10μg\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 298.218px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:10.0000pt\"\u003e100μg\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 298.218px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:11.0000pt\"\u003e\u003cfont\u003eCytokines4\u003c\/font\u003e\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 298.218px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:10.0000pt\"\u003e50ng\/ml\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 298.218px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:10.0000pt\"\u003e5μg\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 298.218px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:10.0000pt\"\u003e50μg\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 298.218px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:11.0000pt\"\u003e\u003cfont\u003eCytokines5\u003c\/font\u003e\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 298.218px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:10.0000pt\"\u003e200ng\/ml\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 298.218px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:10.0000pt\"\u003e20μg\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 298.218px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:10.0000pt\"\u003e200μg\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 298.218px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:11.0000pt\"\u003e\u003cfont\u003eCytokines6\u003c\/font\u003e\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 298.218px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:10.0000pt\"\u003e100ng\/ml\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 298.218px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:10.0000pt\"\u003e10μg\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 298.218px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:10.0000pt\"\u003e100μg\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 298.218px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:11.0000pt\"\u003e\u003cfont\u003eCytokines7\u003c\/font\u003e\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 298.218px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:10.0000pt\"\u003e10ng\/ml\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 298.218px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:10.0000pt\"\u003e5μg\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 298.218px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:10.0000pt\"\u003e10μg\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 298.218px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:11.0000pt\"\u003e\u003cfont\u003eCytokines8\u003c\/font\u003e\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 298.218px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:10.0000pt\"\u003e50ng\/ml\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 298.218px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:10.0000pt\"\u003e5μg\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 298.218px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:10.0000pt\"\u003e50μg\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003c\/tbody\u003e\u003c\/table\u003e\u003c\/div\u003e\u003ch4\u003eProtocol\u003c\/h4\u003e\u003cdiv\u003e\n\u003cp\u003ePreparation of Complete Medium for Mouse Lung Organoids: Combine basal medium, cytokines (1-\u003cfont\u003e8\u003c\/font\u003e), and various additives in specified proportions to prepare the complete medium.\u003c\/p\u003e\n\u003cp\u003e\u003cstrong\u003e1. Primary Culture\u003c\/strong\u003e\u003c\/p\u003e\n\u003cp\u003e(1) Under a biosafety cabinet, extract lung tissue and place it in pre-cooled PBS (supplemented with penicillin, streptomycin, and primary antibiotics) for washing.\u003c\/p\u003e\n\u003cp\u003e(2) Collect the washed tissue fragments into a 1.5 ml EP tube, and mince the tissue as thoroughly as possible (to a homogenate-like consistency) using sterile sharp-pointed ophthalmic scissors. The degree of mincing correlates with digestion time; finer mincing reduces digestion time. Transfer the minced tissue to a 15 ml sterile centrifuge tube.\u003c\/p\u003e\n\u003cp\u003e(3) Add 10 volumes of tissue digestion solution and digest at 37°C with shaking for 10-30 minutes. Monitor the process periodically. Digestion can be terminated when a significant number of cells are observed leaking out under the microscope.\u003c\/p\u003e\n\u003cp\u003e(4) To the confirmed digested tissue suspension, add fetal bovine serum (FBS) to a final concentration of 2-5% or BSA to a final concentration of 0.1%. Mix by pipetting.\u003c\/p\u003e\n\u003cp\u003e(5) Filter the tissue and cell suspension through a 70 µm cell strainer. Centrifuge the filtered cell suspension at 300 g for 3 minutes, then discard the supernatant.\u003c\/p\u003e\n\u003cp\u003e(6) If obvious red precipitate (red blood cells) is observed in the pellet after centrifugation, add 2 ml of red blood cell lysis buffer, mix by pipetting, and incubate for 3 minutes. Centrifuge at 300 g for 3 minutes, then discard the supernatant.\u003c\/p\u003e\n\u003cp\u003e(7) Resuspend the pellet in an appropriate amount of basal medium or PBS.\u003c\/p\u003e\n\u003cp\u003e(8) Mix Matrigel and cells at a suitable ratio. For a 24-well cell culture plate, plate 25-30 µl of the Matrigel-cell mixture per well (perform operations at 4°C).\u003c\/p\u003e\n\u003cp\u003e(9) Place the prepared culture plate in a 37°C incubator for 20-30 minutes for gelation. Add an appropriate amount of complete medium for mouse lung organoids to initiate culture.\u003c\/p\u003e\n\u003cp\u003e\u003cstrong\u003e2. Organoid Passaging\u003c\/strong\u003e\u003c\/p\u003e\n\u003cp\u003e(1) Aspirate the medium using a pipette and add 1-2 ml of 4°C PBS to each well. Incubate for 2 minutes.\u003c\/p\u003e\n\u003cp\u003e(2) Gently disrupt the Matrigel by pipetting and collect the contents into a 15 ml centrifuge tube. Adjust the volume to 10-14 ml with PBS and let it stand at 4°C for 20-30 minutes (grouping every 3-5 wells as a set). Centrifuge at 300 g for 3 minutes, discard the supernatant, and retain the pellet.\u003c\/p\u003e\n\u003cp\u003e(3) Add 1 ml of organoid digestion solution to the collected pellet, mix by pipetting, and incubate at 37°C for 2-3 minutes for digestion. Terminate digestion by adding basal medium. Centrifuge at 300 g for 3 minutes, discard the supernatant, and retain the pellet.\u003c\/p\u003e\n\u003cp\u003e(4) Resuspend the organoids in an appropriate amount of Matrigel. Plate 25-30 µl of the Matrigel-organoid mixture per well in a 24-well cell culture plate. Place the plate in the incubator for 20-30 minutes for gelation, then add an appropriate amount of complete medium for mouse lung organoids to continue culture.\u003c\/p\u003e\n\u003cp\u003e\u003cstrong\u003e3. Organoid Cryopreservation\u003c\/strong\u003e\u003c\/p\u003e\n\u003cp\u003e(1) Aspirate the medium using a pipette and add 1-2 ml of 4°C PBS to each well. Incubate for 2 minutes.\u003c\/p\u003e\n\u003cp\u003e(2) Gently disrupt the Matrigel by pipetting and collect the contents into a 15 ml centrifuge tube. Adjust the volume to 10-14 ml with PBS and let it stand at 4°C for 20-30 minutes (grouping every 3-5 wells as a set). Centrifuge at 300 g for 3 minutes, discard the supernatant, and retain the pellet.\u003c\/p\u003e\n\u003cp\u003e(3) Add an appropriate amount of organoid freezing medium, gently resuspend by pipetting. For a 24-well plate, cryopreserve the pellet from 2-3 wells in one cryovial with a volume of 1 ml per vial.\u003c\/p\u003e\n\u003cp\u003e(4) Label the vials clearly, perform programmed freezing, and then transfer them to liquid nitrogen for long-term storage.\u003c\/p\u003e\n\u003cp\u003e\u003cstrong\u003e4. Organoid Thawing\u003c\/strong\u003e\u003c\/p\u003e\n\u003cp\u003e(1) Pre-aliquot 10 ml of DMEM\/F12 basal medium into a 15 ml centrifuge tube.\u003c\/p\u003e\n\u003cp\u003e(2) Retrieve the cryopreserved organoid vial from the liquid nitrogen tank and quickly thaw it in a 37°C water bath.\u003c\/p\u003e\n\u003cp\u003e(3) During thawing, gently agitate the cryovial to ensure the freezing medium is completely dissolved within 1-2 minutes.\u003c\/p\u003e\n\u003cp\u003e(4) Quickly transfer the thawed organoids to the 15 ml centrifuge tube containing the pre-warmed basal medium. Gently pipette 6-8 times. Centrifuge at 300 g for 3 minutes, then remove the supernatant and collect the organoid pellet.\u003c\/p\u003e\n\u003cp\u003e(5) Resuspend the pellet in Matrigel. Plate 25-30 µl of the Matrigel-organoid mixture per well in a 24-well cell culture plate. Place the plate in the incubator for 20-30 minutes for gelation, then add an appropriate amount of complete medium for mouse lung organoids to initiate culture.\u003cbr\u003e\u003c\/p\u003e\n\u003c\/div\u003e\u003ch4\u003eGuidelines\u003c\/h4\u003e\u003cdiv\u003e\u003cp style=\"margin-bottom: 0px;\"\u003eAvoid vortex oscillation; Store in aliquots to reduce the number of freeze-thaw cycles\u003cbr\u003e\u003c\/p\u003e\u003c\/div\u003e","brand":"UA BIOSCIENCE","offers":[{"title":"100ml","offer_id":41738028056651,"sku":null,"price":800.0,"currency_code":"USD","in_stock":true},{"title":"1000ml","offer_id":41738028089419,"sku":null,"price":4667.0,"currency_code":"USD","in_stock":true}],"thumbnail_url":"\/\/cdn.shopify.com\/s\/files\/1\/0590\/8375\/1499\/files\/2d8ba020-1221-4a85-a78f-09dd04ee1b2d.png?v=1757556265"},{"product_id":"gastric-organoid-cytokine-set-mouse-ua090042","title":"Gastric Organoid Cytokine Set, Mouse","description":"\u003ch4\u003eProduct Specification\u003c\/h4\u003e\u003cdiv class=\"responsive-table product-detail-table details-table\"\u003e\n\u003cbr\u003e\u003ctable style=\"width: 100%; height: auto;\"\u003e\u003ctbody\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 22%;\"\u003e\u003cstrong\u003eSpecies\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 78%;\"\u003eMouse\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 22%;\"\u003e\u003cstrong\u003eEndotoxin\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 78%;\"\u003e\u0026lt;0.1EU\/μg\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 22%;\"\u003e\u003cstrong\u003ePhysical Appearance\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 78%;\"\u003eLyophilized powder\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 22%;\"\u003e\u003cstrong\u003eReconstitution\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 78%;\"\u003e\u003cp style=\"margin-bottom: 0px;\"\u003eReconstitute at 0.1-1 mg\/ml according to the size in ultrapure water after rapid centrifugation.\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 22%;\"\u003e\u003cstrong\u003eStability \u0026amp; Storage\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 78%;\"\u003e\u003cp\u003e·12 months from date of receipt, lyophilized powder stored at -20 to -80℃. \u003cbr\u003e· 3 months, -20 to -80℃ under sterile conditions after reconstitution. \u003cbr\u003e· 1 week, 2 to 8℃ under sterile conditions after reconstitution. \u003cbr\u003e· Please avoid repeated freeze-thaw cycles.\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003c\/tbody\u003e\u003c\/table\u003e\n\u003c\/div\u003e\u003ch4\u003eComponents\u003c\/h4\u003e\u003cdiv\u003e\n\u003ctable class=\"table table-bordered\" style=\"width: 738.41px;\"\u003e\u003ctbody\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 145.669px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:9.0000pt\"\u003eProtein\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 190.009px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:9.0000pt\"\u003eReference dosage\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 186.009px;\"\u003e\u003cp\u003e100 ml system\u003cbr\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 216.002px;\"\u003e\u003cp\u003e1000 ml system\u003cbr\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 298.218px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:9.0000pt\"\u003e\u003cfont\u003e\n\u003cspan style=\"color: rgba(0, 0, 0, 0.9)\"\u003ecytokine\u003c\/span\u003e\n\n\u003c\/font\u003e1\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 298.218px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:9.0000pt\"\u003e100ng\/ml\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 298.218px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:9.0000pt\"\u003e10μg\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 298.218px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:9.0000pt\"\u003e100μg\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 298.218px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:9.0000pt\"\u003e\u003cfont\u003e\n\u003cspan style=\"color: rgba(0, 0, 0, 0.9)\"\u003ecytokine\u003c\/span\u003e\n\n\u003c\/font\u003e2\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 298.218px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:9.0000pt\"\u003e100ng\/ml\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 298.218px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:9.0000pt\"\u003e10μg\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 298.218px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:9.0000pt\"\u003e100μg\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 298.218px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:9.0000pt\"\u003e\u003cfont\u003e\n\u003cspan style=\"color: rgba(0, 0, 0, 0.9)\"\u003ecytokine\u003c\/span\u003e\n\n\u003c\/font\u003e3\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 298.218px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:9.0000pt\"\u003e50ng\/ml\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 298.218px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:9.0000pt\"\u003e5μg\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 298.218px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:9.0000pt\"\u003e50μg\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 298.218px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:9.0000pt\"\u003e\u003cfont\u003e\n\u003cspan style=\"color: rgba(0, 0, 0, 0.9)\"\u003ecytokine\u003c\/span\u003e\n\n\u003c\/font\u003e4\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 298.218px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:9.0000pt\"\u003e100ng\/ml\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 298.218px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:9.0000pt\"\u003e10μg\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 298.218px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:9.0000pt\"\u003e100μg\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 298.218px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:9.0000pt\"\u003e\u003cfont\u003e\n\u003cspan style=\"color: rgba(0, 0, 0, 0.9)\"\u003ecytokine\u003c\/span\u003e\n\n\u003c\/font\u003e5\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 298.218px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:9.0000pt\"\u003e100ng\/ml\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 298.218px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:9.0000pt\"\u003e10μg\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 298.218px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:9.0000pt\"\u003e100μg\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 298.218px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:9.0000pt\"\u003e\u003cfont\u003e\n\u003cspan style=\"color: rgba(0, 0, 0, 0.9)\"\u003ecytokine\u003c\/span\u003e\n\n\u003c\/font\u003e6\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 298.218px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:9.0000pt\"\u003e100ng\/ml\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 298.218px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:9.0000pt\"\u003e10μg\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 298.218px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:9.0000pt\"\u003e100μg\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003c\/tbody\u003e\u003c\/table\u003e\n\u003cp\u003e\u003cbr\u003e\u003c\/p\u003e\n\u003c\/div\u003e\u003ch4\u003eProtocol\u003c\/h4\u003e\u003cdiv\u003e\u003cp\u003ePreparation of Complete Culture Medium for Mouse Gastric Organoids: Mix the basal culture medium, cytokines (1-6), and various additives in the predetermined proportions to prepare the complete culture medium.\u003cbr\u003e\u003cbr\u003e1. Primary Culture\u003cbr\u003e - (1) In the laminar flow cabinet, quickly open the abdominal cavity of the mouse, remove the stomach tissue, and place it in pre - cooled PBS (with penicillin, streptomycin, and primary antibiotics). Open the stomach, rinse the contents, and remove the outer layer of fat and blood vessels.\u003cbr\u003e - (2) Collect the cleaned tissue blocks into a 1.5ml EP tube. Use a sterile fine - tipped ophthalmic surgical scissors to mince the tissue blocks as finely as possible (into a homogenate). The degree of tissue mincing is related to the digestion time. The finer the mincing, the shorter the digestion time. Transfer the minced tissue to a 15ml sterile centrifuge tube.\u003cbr\u003e - (3) Add 10 times the volume of tissue digestion solution for digestion. Incubate at 37℃ with shaking for 10 - 30min. Observe the situation at any time during the digestion process. When a large number of cells are observed to leak out in the field of view and most of the cells are in the form of cell clusters, the digestion can be terminated (note: do not digest to the single - cell state).\u003cbr\u003e - (4) In the tissue suspension confirmed to be digested, add fetal bovine serum (FBS) to a final concentration of 2 - 5% or bovine serum albumin (BSA) to a final concentration of 0.1%, and mix by pipetting.\u003cbr\u003e - (5) Pass the tissue and cell suspension through a 70µm cell strainer. Centrifuge the filtered cell suspension at 1000rpm for 5min. After centrifugation, discard the supernatant.\u003cbr\u003e - (6) If obvious red precipitate (red blood cells) is observed in the precipitate after centrifugation, add 2ml of red blood cell lysis solution, mix by pipetting, and leave it for 3min. Centrifuge at 1000rpm for 5min, and then discard the supernatant.\u003cbr\u003e - (7) Resuspend the precipitate with an appropriate amount of basic culture medium or PBS.\u003cbr\u003e - (8) Mix the matrix gel and crypts in a suitable proportion. For example, for a 24 - well cell culture plate, spot 25 - 30ul of the matrix gel mixture into each well for coating (operate at 4℃).\u003cbr\u003e - (9) Place the coated culture plate in a 37℃ incubator for 20 - 30min to gel. Add an appropriate amount of mouse stomach organoid complete culture medium (at room temperature) for culture.\u003cbr\u003e\u003cbr\u003e2. Organoid Passaging Culture\u003cbr\u003e - (1) Aspirate the culture medium with a pipette, add 1 - 2ml of 4℃ PBS to each well, and leave it for 2min.\u003cbr\u003e - (2) Gently pipette the matrix gel to collect it in a 15ml centrifuge tube. Adjust the volume with PBS to 10 - 14ml, and leave it at 4℃ for 20 - 30min. Centrifuge at 1000rpm for 5min to discard the liquid and retain the precipitate (group every 3 - 5 wells).\u003cbr\u003e - (3) Resuspend the organoids with an appropriate amount of matrix gel, and spot 25 - 30ul of matrix gel into each well of a 24 - well cell culture plate. Place it in the incubator for 20 - 30min to gel, and then add an appropriate amount of mouse stomach organoid complete culture medium.\u003cbr\u003e\u003cbr\u003e3. Organoid Cryopreservation\u003cbr\u003e - (1) Aspirate the culture medium with a pipette, add 1 - 2ml of 4℃ PBS to each well, and leave it for 2min.\u003cbr\u003e - (2) Gently pipette the matrix gel to collect it in a 15ml centrifuge tube. Adjust the volume with PBS to 10 - 14ml, and leave it at 4℃ for 20 - 30min. Centrifuge at 1000rpm for 5min to discard the liquid and retain the precipitate (group every 3 - 5 wells).\u003cbr\u003e - (3) Add an appropriate amount of organoid cryopreservation solution, and gently pipette to resuspend. For example, for a 24 - well cell culture plate, cryopreserve 2 - 3 wells in one tube, with a volume of 1ml per tube.\u003cbr\u003e - (4) Label the information properly, and then transfer it to liquid nitrogen for long - term storage after programmed cooling.\u003cbr\u003e\u003cbr\u003e4. Organoid Thawing\u003cbr\u003e - (1) Take 10ml of DMEM\/F12 basic culture medium in a 15ml centrifuge tube.\u003cbr\u003e - (2) Remove the cryopreserved organoids from the liquid nitrogen tank, and quickly place them in a 37℃ water bath to thaw.\u003cbr\u003e - (3) During the water bath thawing process, gently shake the cryovial to ensure that the cryopreservation solution is completely thawed within 1 - 2min.\u003cbr\u003e - (4) Quickly transfer the thawed organoids to a 15ml centrifuge tube, gently pipette 6 - 8 times with a pipette, centrifuge at 1000rpm for 5min, and then remove the supernatant to collect the organoid precipitate.\u003cbr\u003e - (5) Resuspend with matrix gel, spot 25 - 30ul of matrix gel into each well of a 24 - well cell culture plate, place it in the incubator for 20 - 30min to gel, and then add an appropriate amount of mouse stomach organoid complete culture medium.\u003cbr\u003e\u003c\/p\u003e\u003c\/div\u003e\u003ch4\u003eGuidelines\u003c\/h4\u003e\u003cdiv\u003e\u003cp style=\"margin-bottom: 0px;\"\u003eAvoid vortex oscillation; Store in aliquots to reduce the number of freeze-thaw cycles\u003cbr\u003e\u003c\/p\u003e\u003c\/div\u003e","brand":"UA BIOSCIENCE","offers":[{"title":"100ml","offer_id":41738033758283,"sku":null,"price":696.0,"currency_code":"USD","in_stock":true},{"title":"1000ml","offer_id":41738033791051,"sku":null,"price":3180.0,"currency_code":"USD","in_stock":true}],"thumbnail_url":"\/\/cdn.shopify.com\/s\/files\/1\/0590\/8375\/1499\/files\/AntBioImage_3cbe4c37-373e-4c0e-8f87-956ef7bbe6a1.png?v=1770719289"},{"product_id":"cholangiocyte-organoid-expansion-cytokine-set-mouse-ua090044","title":"Cholangiocyte Organoid Expansion Cytokine Set, Mouse","description":"\u003ch4\u003eProduct Specification\u003c\/h4\u003e\u003cdiv class=\"responsive-table product-detail-table details-table\"\u003e\n\u003cbr\u003e\u003ctable style=\"width: 100%; height: auto;\"\u003e\u003ctbody\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 22%;\"\u003e\u003cstrong\u003eSpecies\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 78%;\"\u003eMouse\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 22%;\"\u003e\u003cstrong\u003eEndotoxin\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 78%;\"\u003e\u0026lt;0.1EU\/μg\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 22%;\"\u003e\u003cstrong\u003ePhysical Appearance\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 78%;\"\u003eLyophilized powder\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 22%;\"\u003e\u003cstrong\u003eReconstitution\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 78%;\"\u003e\u003cp style=\"margin-bottom: 0px;\"\u003eReconstitute at 0.1-1 mg\/ml according to the size in ultrapure water after rapid centrifugation.\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 22%;\"\u003e\u003cstrong\u003eStability \u0026amp; Storage\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 78%;\"\u003e\u003cp\u003e·12 months from date of receipt, lyophilized powder stored at -20 to -80℃. \u003cbr\u003e· 3 months, -20 to -80℃ under sterile conditions after reconstitution. \u003cbr\u003e· 1 week, 2 to 8℃ under sterile conditions after reconstitution. \u003cbr\u003e· Please avoid repeated freeze-thaw cycles.\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003c\/tbody\u003e\u003c\/table\u003e\n\u003c\/div\u003e\u003ch4\u003eComponents\u003c\/h4\u003e\u003cdiv\u003e\n\u003ctable class=\"table table-bordered\" style=\"width: 638.41px;\"\u003e\u003ctbody\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 133.669px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:9.0000pt\"\u003eProtein\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 204.012px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:9.0000pt\"\u003eReference dosage\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 152.007px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:9.0000pt\"\u003e100ml system\u003cfont\u003e\u003c\/font\u003e\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 148.005px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:9.0000pt\"\u003e1000m system\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 298.218px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:9.0000pt\"\u003e\u003cfont\u003eCytokines \u003c\/font\u003e1\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 298.218px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:9.0000pt\"\u003e50ng\/ml\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 298.218px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:9.0000pt\"\u003e5μg\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 298.218px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:9.0000pt\"\u003e50μg\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 298.218px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:9.0000pt\"\u003e\u003cfont\u003eCytokines \u003c\/font\u003e2\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 298.218px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:9.0000pt\"\u003e250ng\/ml\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 298.218px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:9.0000pt\"\u003e25μg\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 298.218px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:9.0000pt\"\u003e250μg\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 298.218px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:9.0000pt\"\u003e\u003cfont\u003eCytokines \u003c\/font\u003e3\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 298.218px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:9.0000pt\"\u003e50ng\/ml\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 298.218px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:9.0000pt\"\u003e5μg\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 298.218px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:9.0000pt\"\u003e50μg\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 298.218px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:9.0000pt\"\u003e\u003cfont\u003eCytokines \u003c\/font\u003e4\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 298.218px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:9.0000pt\"\u003e100ng\/ml\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 298.218px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:9.0000pt\"\u003e10μg\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 298.218px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:9.0000pt\"\u003e100μg\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 298.218px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:9.0000pt\"\u003e\u003cfont\u003eCytokines \u003c\/font\u003e5\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 298.218px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:9.0000pt\"\u003e25ng\/ml\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 298.218px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:9.0000pt\"\u003e5μg\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 298.218px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:9.0000pt\"\u003e25μg\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003c\/tbody\u003e\u003c\/table\u003e\n\u003cp\u003e\u003cbr\u003e\u003c\/p\u003e\n\u003c\/div\u003e\u003ch4\u003eProtocol\u003c\/h4\u003e\u003cdiv\u003e\n\u003cp\u003ePreparation of Complete Medium (Expansion) for Mouse Cholangiocyte Organoids:\u003c\/p\u003e\n\u003cp\u003eMix the basal medium, cytokines (1-\u003cfont\u003e5\u003c\/font\u003e), and various supplements in specified proportions to prepare the complete medium.\u003c\/p\u003e\n\u003cp\u003e1. Primary Culture\u003c\/p\u003e\n\u003cp\u003e(1) Quickly dissect the abdominal cavity of the mouse in a biosafety cabinet, extract the liver tissue, and rinse it in pre-cooled PBS (supplemented with penicillin, streptomycin, and primary antibiotics).\u003c\/p\u003e\n\u003cp\u003e(2) Collect the rinsed tissue fragments into a 1.5 ml EP tube, and mince the tissue as thoroughly as possible (to a homogenate-like consistency) using sterile sharp-tipped ophthalmic surgical scissors. The degree of mincing affects digestion time; finer mincing shortens digestion time. Transfer the minced tissue to a 15 ml sterile centrifuge tube.\u003c\/p\u003e\n\u003cp\u003e(3) Add 10 volumes of tissue digestion solution and digest at 37°C with shaking for 10–30 minutes. Monitor the process closely. Digestion can be terminated when a large number of cells are released and most cells appear as clusters (Note: Avoid digesting into a single-cell state).\u003c\/p\u003e\n\u003cp\u003e(4) After confirming digestion, add fetal bovine serum (FBS) to a final concentration of 2–5% or BSA to a final concentration of 0.1%, and mix by pipetting.\u003c\/p\u003e\n\u003cp\u003e(5) Filter the tissue and cell suspension through a 70 µm cell strainer. Centrifuge the filtered cell suspension at 1000 rpm for 5 minutes, and discard the supernatant.\u003c\/p\u003e\n\u003cp\u003e(6) If obvious red precipitate (red blood cells) is observed in the pellet after centrifugation, add 2 ml of red blood cell lysis buffer, mix by pipetting, and incubate for 3 minutes. Centrifuge at 1000 rpm for 5 minutes and discard the supernatant.\u003c\/p\u003e\n\u003cp\u003e(7) Resuspend the pellet in an appropriate amount of basal medium or PBS.\u003c\/p\u003e\n\u003cp\u003e(8) Mix the basement membrane matrix (e.g., Matrigel) and cells at a suitable ratio. For a 24-well cell culture plate, plate 25–30 µl of the matrix-cell mixture per well (perform\u003cfont\u003e操作 at 4°C).\u003c\/font\u003e\u003c\/p\u003e\n\u003cp\u003e(9) Place the plate in a 37°C incubator for 20–30 minutes to allow gelation. Add an appropriate amount of complete medium (expansion) for mouse cholangiocyte organoids for culture.\u003c\/p\u003e\n\u003cp\u003e2. Passaging of Organoids\u003c\/p\u003e\n\u003cp\u003e(1) Aspirate the medium using a pipette, add 1–2 ml of 4°C PBS per well, and incubate for 2 minutes.\u003c\/p\u003e\n\u003cp\u003e(2) Gently pipette to disrupt the matrix, collect the contents into a 15 ml centrifuge tube, adjust the volume to 10–14 ml with PBS, and incubate at 4°C for 20–30 minutes (group every 3–5 wells together). Centrifuge at 1000 rpm for 5 minutes, discard the supernatant, and retain the pellet.\u003c\/p\u003e\n\u003cp\u003e(3) Resuspend the organoids in an appropriate amount of matrix, plate 25–30 µl per well in a 24-well cell culture plate, and incubate for 20–30 minutes to allow gelation. Add an appropriate amount of complete medium (expansion) for mouse cholangiocyte organoids for culture.\u003c\/p\u003e\n\u003cp\u003e3. Cryopreservation of Organoids\u003c\/p\u003e\n\u003cp\u003e(1) Aspirate the medium using a pipette, add 1–2 ml of 4°C PBS per well, and incubate for 2 minutes.\u003c\/p\u003e\n\u003cp\u003e(2) Gently pipette to disrupt the matrix, collect the contents into a 15 ml centrifuge tube, adjust the volume to 10–14 ml with PBS, and incubate at 4°C for 20–30 minutes (group every 3–5 wells together). Centrifuge at 1000 rpm for 5 minutes, discard the supernatant, and retain the pellet.\u003c\/p\u003e\n\u003cp\u003e(3) Add an appropriate amount of organoid cryopreservation medium, gently resuspend by pipetting, and aliquot. For a 24-well plate, cryopreserve the equivalent of 2–3 wells per cryovial, with 1 ml per vial.\u003c\/p\u003e\n\u003cp\u003e(4) Label the vials appropriately, perform programmed freezing, and transfer to liquid nitrogen for long-term storage.\u003c\/p\u003e\n\u003cp\u003e4. Thawing of Organoids\u003c\/p\u003e\n\u003cp\u003e(1) Pre-aliquot 10 ml of DMEM\/F12 basal medium into a 15 ml centrifuge tube.\u003c\/p\u003e\n\u003cp\u003e(2) Retrieve the cryopreserved organoids from the liquid nitrogen tank and quickly thaw them in a 37°C water bath.\u003c\/p\u003e\n\u003cp\u003e(3) During thawing, gently swirl the cryovial to ensure complete dissolution of the cryopreservation medium within 1–2 minutes.\u003c\/p\u003e\n\u003cp\u003e(4) Quickly transfer the thawed organoids to the 15 ml centrifuge tube, gently pipette 6–8 times, centrifuge at 1000 rpm for 5 minutes, then discard the supernatant and collect the organoid pellet.\u003c\/p\u003e\n\u003cp\u003e(5) Resuspend in matrix, plate 25–30 µl per well in a 24-well cell culture plate, and incubate for 20–30 minutes to allow gelation. Add an appropriate amount of complete medium (expansion) for mouse cholangiocyte organoids for culture.\u003cbr\u003e\u003c\/p\u003e\n\u003c\/div\u003e\u003ch4\u003eGuidelines\u003c\/h4\u003e\u003cdiv\u003e\u003cp style=\"margin-bottom: 0px;\"\u003eAvoid vortex oscillation; store in separate containers to reduce the number of freeze-thaw cycles.\u003c\/p\u003e\u003c\/div\u003e","brand":"UA BIOSCIENCE","offers":[{"title":"100ml","offer_id":41738036314187,"sku":null,"price":596.0,"currency_code":"USD","in_stock":true},{"title":"1000ml","offer_id":41738036346955,"sku":null,"price":2646.0,"currency_code":"USD","in_stock":true}],"thumbnail_url":"\/\/cdn.shopify.com\/s\/files\/1\/0590\/8375\/1499\/files\/AntBioImage_38ad7bd2-e576-4d61-b557-fbc1d5306bab.png?v=1770719277"}],"url":"https:\/\/www.antbioinc.com\/collections\/3d-organoid-culture-1-2.oembed","provider":"AntBio","version":"1.0","type":"link"}