{"product_id":"parathyroid-gland-organoid-culture-kit-abs90627","title":"Parathyroid gland Organoid Culture Kit","description":"\u003ch4\u003eProduct Specification\u003c\/h4\u003e\u003cdiv class=\"responsive-table product-detail-table details-table\"\u003e\u003ctable\u003e\u003ctbody\u003e\n\u003ctr\u003e\n\u003ctd\u003e\u003cstrong\u003eUsage\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd\u003e\n\u003cp\u003e\u003cstrong\u003e I. Pre-Organization Processing \u003cbr\u003e\u003c\/strong\u003e\u003cstrong\u003e1 Experimental materials \u003cbr\u003e\u003c\/strong\u003e Primary buffer needs to be prepared in advance B （ 4 ℃ Pre-cooling), tissue preservation solution E Sampling tube, tissue transport box, ice pack. \u003cbr\u003e\u003cstrong\u003e2 Organization acquisition and transportation \u003cbr\u003e\u003c\/strong\u003e Tissue sampling and transportation are the first and most easily overlooked steps in the successful construction of organoids. Improper preservation of tissues in the early stages can lead to problems such as poor cell activity, contamination, and few normal cells, reducing the success rate of organoid construction. \u003cbr\u003e The tissue should be in vitro 30 min Place tissue preservation solution inside E , primary buffer B cleaning 3-5 Next, the blood on the tissue surface is washed clean and placed in tissue preservation solution. E In the sampling tube 4 ℃ Cryogenic storage and transportation ( 72 h inside). \u003cbr\u003e\u003cstrong\u003e\u003cbr\u003e II. Primary organoid culture (with 24 (Taking orifice plates as an example) \u003cbr\u003e\u003c\/strong\u003e\u003cstrong\u003e1 、 \u003c\/strong\u003e\u003cstrong\u003e experimental material \u003cbr\u003e\u003c\/strong\u003e Primary buffer needs to be prepared in advance B （ 4 ℃ ), primary tissue digestive juice C （ 37 ℃ ), matrix glue (in advance 24 h Put in 4 ℃ (Refrigerator melting)   thyroid organoid medium A (Room temperature or 37 ℃ ), tweezers ( 10㎝ ), pointed ophthalmic surgical scissors \/ Surgical blade, disposable 60 mm Petri dishes, 1.5 mL\/15 mL\/50 mL centrifuge tube, 100㎛ a cell screen, 3 mL pasteurized straw \/1000 ul pipetting gun, 24 Well cell culture plate, metal ice box. \u003cbr\u003e\u003cstrong\u003e2 Construction of parathyroid organoids \u003cbr\u003e\u003c\/strong\u003e\u003cstrong\u003e2.1 Organizational processing \u003cbr\u003e\u003c\/strong\u003e It is recommended that the parathyroid organoid tissue after sampling be  2 – 8 ℃ Store and transport under these conditions, and quickly transfer to a clean laboratory for thyroid organoid construction experiments. Organize photography and record detailed information. \u003cbr\u003e\u003cstrong\u003e2.1.1 Tissue cleansing \u003cbr\u003e\u003c\/strong\u003e After the sampling tube is sterilized, the tissue is removed from the ultra-clean table, placed in a petri dish, and primary buffer is added.  B , use 3 mL pasteurized straw or 1000 ul The pipette gun is blown and cleaned, and the cleaning operation is repeated three or more times. \u003cbr\u003e\u003cstrong\u003e2.1.2 Tissue dissociation and digestion \u003c\/strong\u003e\u003cstrong\u003e \u003cbr\u003e\u003c\/strong\u003e Tissue impurities are removed with ophthalmic scissors or a surgical blade, and the forceps are transferred to 1.5 mL EP In the tube, ophthalmic scissors are used to further mechanically dissociate the tissue into a volume of approximately  1~3 mm\u003cstrong\u003e\u003csup\u003e3\u003c\/sup\u003e\u003c\/strong\u003e The tissue block was transferred to 15 mL EP In the tube, add 5 mL primary tissue digestive fluid 37 ℃ oscillation digestion 15-25 min 。 Each digestive process 10 min The tissue digestion was observed under a microscope. A small amount of digestive juice was taken and observed under the microscope, and a larger amount was observed. 70 μm The following cell clusters or individual cells are followed by the next step. The degree of tissue digestion is shown in Figure 1. \u003cbr\u003e\u003cstrong\u003e If the amount of tissue is too small or the biopsy tissue is used \u003c\/strong\u003e\u003cstrong\u003e1 mL primary tissue digestive fluid C In 1.5 mL EP Tubular digestion. \u003c\/strong\u003e\u003c\/p\u003e\n\u003cdiv style=\"text-align: center;\"\u003e\n\u003cimg src=\"https:\/\/absin.oss-cn-shanghai.aliyuncs.com\/absin-china\/20260402\/46cff56ea1614009a12743f92a5dff52.jpg\" alt=\"\" width=\"454\" height=\"200\"\u003e\u003cbr\u003e Figure 1   Tissue digestion into more cell clusters or more single cells \u003c\/div\u003e\n\u003cp\u003e\u003cstrong\u003e2.1.3 、 \u003c\/strong\u003e\u003cstrong\u003e Filtration of tissue \u003cbr\u003e\u003c\/strong\u003e After digestion is complete, the tissue vortex is scrolled. 10s , join 3 Double the volume of primary buffer B Digestion is terminated, and the digestive tissue mixture passes through. 100 μm Cell sieve filtration with pore size, continue with 10-20ml primary buffer B Rinsing the tissue to collect more tissue mass cells, 300 g enrichment centrifugation 5 min Abandoning the supernatural; use 8-10ml primary buffer B Resuspend the centrifugal precipitate (remove more impurities), 300 g enrichment centrifugation 5 min They abandoned their superiors. \u003cbr\u003e If the cell pellet contains red blood cells, add 1-2 mL erythrocyte lysate 1-2 min Then, dilute to 10 mL ， 300 g enrichment centrifugation 5 min They abandoned their superiors. \u003cstrong\u003e Organoid culture was performed directly when there was too little sedimentation or no red blood cells. \u003cbr\u003e\u003cbr\u003e\u003c\/strong\u003e\u003cstrong\u003e2.1.4 Tumor organoid culture \u003cbr\u003e\u003c\/strong\u003e Observe the volume of cell pellets collected by centrifugation, and add 25 Double the volume of matrix glue is resuspended, forming 3D Cultivate the spatial structure to avoid bubbles during the resuspension process. The cell pellet volume is shown in Figure 2. Add the cell pellet volume according to the figure. 200 ul 、 150 ul 、 100 ul 、 50 ul Matrix glue. \u003cbr\u003e\u003cimg style=\"display: block; margin-left: auto; margin-right: auto;\" src=\"https:\/\/absin.oss-cn-shanghai.aliyuncs.com\/absin-china\/20260402\/d63bbeb8ce91434998e218fce04ebc79.png\" alt=\"\" width=\"339\" height=\"153\"\u003e                                                                                                         Figure 2   cell pellet volume \u003c\/p\u003e\n\u003cp\u003e24 well cell culture plate according to 25 ul-30 ul\/ When dispensing the adhesive through the holes, the matrix adhesive is maintained throughout the entire process. 0-4℃ Operate under these conditions. cell culture plate placement 37 ℃ Incubator 10-15 min After the matrix gum has set, add to each well. 500-750 ul  Parathyroid organoid medium A (in advance) 37 ℃ (Preheating) for cultivation. \u003cstrong\u003e\u003cbr\u003e\u003cbr\u003e\u003cstrong\u003e III. \u003c\/strong\u003e\u003cstrong\u003e Organoid subculture (with 24 (Taking orifice plates as an example) \u003c\/strong\u003e\u003c\/strong\u003e\u003c\/p\u003e\n\u003cp\u003e\u003cstrong\u003e1 Experimental materials \u003c\/strong\u003e\u003c\/p\u003e\n\u003cp\u003e\u003cstrong\u003e passage buffer G （ 4 ℃ ), organoid digestive fluid D (Room temperature or 37 ℃ ), matrix glue (in advance 24 h Put in 4 ℃ (Refrigerator melting)   Parathyroid organoid medium A (Room temperature or 37 ℃ ）、 1.5 mL\/15 mL centrifuge tube, 24 Well cell culture plate, ice box. \u003cbr\u003e\u003c\/strong\u003e\u003c\/p\u003e\n\u003cp\u003e\u003cstrong\u003e2 、 \u003c\/strong\u003e\u003cstrong\u003e organoid passage \u003c\/strong\u003e\u003c\/p\u003e\n\u003cp\u003e Select suitable organoids for passage, generally after about a week of growth, microscope 10X You can see more below 20 Organoids, or size 100-200μm Organoids. \u003cbr\u003e Aspirate off the medium and add an equal volume of passage buffer to each well G The pipette gently blows away the matrix glue, collecting it from... 15 mL In the centrifuge tube, each 6-8 The hole is transferred to a centrifuge tube, 4 ℃ standing 10-15 min 。 \u003cbr\u003e\u003cbr\u003e\u003cstrong\u003e2.1 Organoid digestion \u003cbr\u003e\u003c\/strong\u003e The need for digestive passaging is determined based on the growth of the organoid. \u003cstrong\u003e After centrifugation, if there is little precipitation at the bottom of the tube, no cells are found, and the matrix glue is not delaminated, it can be resuspended again to increase the centrifugal force and centrifuge again. \u003c\/strong\u003e\u003cstrong\u003e 。 \u003cbr\u003e\u003c\/strong\u003e When the number of organoids is insufficient or the volume is small, 300 g centrifuge 5 min Abandon the Shangqing. \u003cbr\u003e When the number of organoids is high or the volume is large, 300 g centrifuge 5 min Discard the supernatant, and you can choose digestive juice digestion or mechanical digestion. \u003cbr\u003e Digestive juice digestion: Add 1-2 mL organoid digestive fluid D After the cell pellet is blown out, it is digested at room temperature. 2-4 min Blow it every minute, each time... 20 Observe under a microscope until digestion reaches (Figure 3). A-B It can be stopped when in the ) state. Add 3 Passage buffer with twice the volume of organoid digestive fluid G Termination of digestion, 300 g centrifuge 5 min They abandoned the Shangqing \u003cbr\u003e Mechanical digestion: Add 1 mL passage buffer G ， 1 mL gun muzzle blowing 20-30 Next, until the blowing and beating reach (Figure 3). C-D It can be stopped and added when in the ) state. 10 mL passage buffer, 300 g centrifuge 5 min 。 \u003cimg style=\"display: block; margin-left: auto; margin-right: auto;\" src=\"https:\/\/absin.oss-cn-shanghai.aliyuncs.com\/absin-china\/20260402\/b64e8b46d88d42119f7716ea09f7fdb4.jpg\" alt=\"\" width=\"635\" height=\"288\"\u003e                                                                        Figure 3   Organoid passage digestion degree diagram \u003c\/p\u003e\n\u003cp\u003e\u003cstrong\u003e2.2 Passage organoid culture \u003cbr\u003e\u003c\/strong\u003e Observe the volume of organoid pellets collected by centrifugation; if there is very little pellet, it can be reserved. 1 Dispense the supernatant with twice the precipitation volume; a large amount of precipitated supernatant can be absorbed. Add... 25 The organoids were resuspended in an amount of matrix gel twice the organoid pellet volume. The volume of matrix rubber can be referenced. “ Organoid primary culture operation Figure 2 ” 。 \u003cbr\u003e24 well cell culture plate according to 25ul-30ul\/ Dispensing adhesive through holes \u003cstrong\u003e The matrix adhesive is maintained throughout the 0-4 ℃ operation under conditions \u003c\/strong\u003e 。 cell culture plate placement 37 ℃ Incubator 10-15 min After the matrix gum has set, add to each well. 500-750 ul  Parathyroid organoid medium A (in advance) 37 ℃ (Preheating) for cultivation. \u003cbr\u003e\u003cstrong\u003e\u003cbr\u003e IV. \u003c\/strong\u003e\u003cstrong\u003e Cryopreservation of organoids (with 24 (Taking orifice plates as an example) \u003cbr\u003e\u003c\/strong\u003e\u003cstrong\u003e1 Experimental materials \u003cbr\u003e\u003c\/strong\u003e passage buffer G （ 4 ℃ ), organoid cryopreservation solution F （ 4 ℃ ）、 15 mL Centrifuge tube, cell cryopreservation tube, program cooling box, pipette gun. \u003cbr\u003e\u003cstrong\u003e2 Organoid cryopreservation \u003cbr\u003e\u003c\/strong\u003e Organoids that are temporarily unused should be frozen and stored in a low-temperature environment. \u003cbr\u003e Aspirate off the medium and add an equal volume of passage buffer to each well G The pipette gently blows away the matrix glue, collecting it from... 15 mL In the centrifuge tube, each 6-8 The hole is transferred to a centrifuge tube, 4 ℃ standing 10-15 min 。 300 g centrifuge 5 min Discard the supernatant and add it every three holes. 2 mL organoid cryopreservation fluid F Gently blow and mix well, then transfer to cell cryopreservation tubes. Each tube... 1 mL 。 \u003cbr\u003e Mark the information, place it in the program cooling box, and move it to... -80 ℃ In the refrigerator, 48 h Afterwards, it was placed in a liquid nitrogen tank for storage. Or put in 4 ℃ refrigerator 40 min Then, put in -20 ℃ in the refrigerator 2 h , move to -80 ℃ In the refrigerator, 48 h Afterwards, it was placed in a liquid nitrogen tank for storage. \u003cbr\u003e\u003cbr\u003e\u003cstrong\u003e V. \u003c\/strong\u003e\u003cstrong\u003e organoid resuscitation \u003c\/strong\u003e\u003cstrong\u003e cultivate \u003c\/strong\u003e\u003cstrong\u003e (with 24 (Taking orifice plates as an example) \u003c\/strong\u003e\u003c\/p\u003e\n\u003cp\u003e\u003cstrong\u003e1 Experimental materials \u003cbr\u003e\u003c\/strong\u003e passage buffer G 、   thyroid organoid medium A Matrix glue (in advance) 24 h Put in 4 ℃ (Refrigerator melting) 24 well cell culture plates, ice boxes, 15mL Centrifuge tube, water bath, 3mL pasteurized straw \/ Pipetting gun. \u003cbr\u003e\u003cstrong\u003e2 Organoid resuscitation culture \u003cbr\u003e\u003c\/strong\u003e （ 1 The frozen organoids were removed from the cryogenic environment and quickly placed in... 37 ℃ Thawing in a water bath. During the water bath thawing process, the cryopreservation tube needs to be gently shaken to ensure that the cryopreservation liquid is completely thawed in a short time. \u003cbr\u003e （ 2 During the organoid dissolution process, to 15 mL Advance addition to the centrifuge tube 10 mL Passage buffer for backup; \u003cbr\u003e （ 3 ) Use 1 mL Gently pipette the cryopreserved fluid, slowly add it to the prepared buffer, and gently blow with the pipette. 2-3 next time, 300 g centrifuge 5 min Abandon the Shangqing; \u003cbr\u003e （ 4 Add approximately [amount missing] to the 10 mL Passage buffer, gently blown with a pipette 2-3 Next (the precipitate can be blown away), 300 g centrifuge 5 min Abandon the Shangqing; \u003cbr\u003e （ 5 ) Use 200 μL Carefully pipette the residual supernatant and add... 200-400 μL Matrix glue (usual: resuscitation of one tube of cryopreserved organoids) 200 μL Matrix glue, resuscitation 2 Use of organoids cryopreserved in tubes 400 μL The matrix glue is resuspended (to avoid bubbles). 24 well cell culture plate according to 25 μL-30 μL Adhesive is dispensed per well, and the matrix adhesive is maintained at the same level 0-4 ℃ Operate under conditions; \u003cbr\u003e （ 6 ) Cell culture plate placement 37 ℃ Carbon dioxide incubator 10-15 min After the matrix gum has set, add to each well. 500-750 μL Organoid medium (in advance) 37 ℃ (Preheating) for cultivation. \u003cbr\u003e\u003cbr\u003e\u003cstrong\u003e VI. \u003c\/strong\u003e\u003cstrong\u003e Matrix rubber use \u003cbr\u003e\u003c\/strong\u003e In 2–8 ℃ The matrix glue was thawed overnight under ambient conditions. When using matrix glue, keep it in an ice box to prevent premature setting. Matrix glue in 37 ℃ Below 20 A gel formed within minutes. \u003cbr\u003e\u003cstrong\u003e1 Matrix glue \u003c\/strong\u003e\u003cstrong\u003e Features: \u003cbr\u003e\u003c\/strong\u003e4℃ continuous 14 Good liquidity can still be maintained today. \u003cbr\u003e Put in 37℃ Incubator 10-15 min It can solidify \u003cbr\u003e It is not easily damaged during the culture process, and the culture plate is clean and does not stick when the glue is removed. \u003c\/p\u003e\n\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003e\u003cstrong\u003eDescription\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd\u003e\n\u003cstrong\u003e\u003cstrong\u003e Kit Composition: \u003cbr\u003e\u003c\/strong\u003e\u003c\/strong\u003e\u003ctable style=\"border-collapse: collapse; width: 95%; height: 169px; margin-left: auto; margin-right: auto;\" border=\"1\"\u003e\u003ctbody\u003e\n\u003ctr style=\"height: 22px;\"\u003e\n\u003ctd style=\"width: 22.5414%; text-align: center; height: 22px;\"\u003e Component Name \u003c\/td\u003e\n\u003ctd style=\"width: 13.5761%; text-align: center; height: 22px;\"\u003e Specification \u003c\/td\u003e\n\u003ctd style=\"width: 22.6256%; text-align: center;\"\u003e Storage conditions \u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr style=\"height: 22px;\"\u003e\n\u003ctd style=\"width: 22.5414%; height: 22px;\"\u003e parathyroid organoid culture medium A\u003c\/td\u003e\n\u003ctd style=\"width: 13.5761%; height: 22px;\"\u003e100mL\u003c\/td\u003e\n\u003ctd style=\"width: 22.6256%;\"\u003e4 ℃ ，  3 months or -20 ℃ ， 1 year\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr style=\"height: 22px;\"\u003e\n\u003ctd style=\"width: 22.5414%; height: 22px;\"\u003e primary buffer  B\u003c\/td\u003e\n\u003ctd style=\"width: 13.5761%; height: 22px;\"\u003e250mL\u003c\/td\u003e\n\u003ctd style=\"width: 22.6256%;\"\u003e4\/-20 ℃ ， 2 year\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr style=\"height: 22px;\"\u003e\n\u003ctd style=\"width: 22.5414%; height: 22px;\"\u003e primary tissue digestive fluid C\u003c\/td\u003e\n\u003ctd style=\"width: 13.5761%; height: 22px;\"\u003e30mL\u003c\/td\u003e\n\u003ctd style=\"width: 22.6256%;\"\u003e4\/-20 ℃ ， 1 year\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr style=\"height: 22px;\"\u003e\n\u003ctd style=\"width: 22.5414%; height: 22px;\"\u003e organoid digestive fluid  D\u003c\/td\u003e\n\u003ctd style=\"width: 13.5761%; height: 22px;\"\u003e30mL\u003c\/td\u003e\n\u003ctd style=\"width: 22.6256%;\"\u003e4\/-20 ℃ ， 1 year\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr style=\"height: 22px;\"\u003e\n\u003ctd style=\"width: 22.5414%; height: 22px;\"\u003e tissue preservation solution  E\u003c\/td\u003e\n\u003ctd style=\"width: 13.5761%; height: 22px;\"\u003e100mL\u003c\/td\u003e\n\u003ctd style=\"width: 22.6256%;\"\u003e4\/-20 ℃ ， 1 year\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr style=\"height: 22px;\"\u003e\n\u003ctd style=\"width: 22.5414%; height: 22px;\"\u003e organoid cryopreservation fluid  F\u003c\/td\u003e\n\u003ctd style=\"width: 13.5761%; height: 22px;\"\u003e20mL\u003c\/td\u003e\n\u003ctd style=\"width: 22.6256%;\"\u003e4\/-20 ℃ ， 2 year\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr style=\"height: 15px;\"\u003e\n\u003ctd style=\"width: 22.5414%; height: 15px;\"\u003e passage buffer  G\u003c\/td\u003e\n\u003ctd style=\"width: 13.5761%; height: 15px;\"\u003e250mL\u003c\/td\u003e\n\u003ctd style=\"width: 22.6256%;\"\u003e4\/-20 ℃ ， 2 year\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003c\/tbody\u003e\u003c\/table\u003e\n\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003e\u003cstrong\u003eGeneral Notes\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd\u003e\n\u003cp\u003e\u003cstrong\u003e1. Parathyroid organoid culture medium\u003c\/strong\u003eA can be stored at 4℃ for 3 months. After receiving the goods, store them at 4℃. It is recommended to use them within 1 month. If not used for a long time, it is recommended to store them at -20℃ to avoid freezing and thawing more than twice.\u003cstrong\u003e。\u003cbr\u003e2. Tissue preservation solution\u003c\/strong\u003eand\u003cstrong\u003ePrimary tissue digestive juice\u003c\/strong\u003eC contains nutrients that maintain cell activity. To maintain the activity of the reagent nutrients, it is recommended to store it at -20℃ without long-term use to avoid repeated freezing and thawing more than twice.\u003cstrong\u003e。\u003c\/strong\u003e\u003c\/p\u003e\n\u003cp\u003e\u003cstrong\u003e \u003c\/strong\u003e\u003c\/p\u003e\n\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003e\u003cstrong\u003eStorage Temp.\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd\u003eStore at -20℃ and shelf life is one year.\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003c\/tbody\u003e\u003c\/table\u003e\u003c\/div\u003e","brand":"Absin","offers":[{"title":"1Kit","offer_id":43224466391115,"sku":"abs90627-1Kit","price":873.0,"currency_code":"USD","in_stock":true}],"url":"https:\/\/www.antbioinc.com\/products\/parathyroid-gland-organoid-culture-kit-abs90627","provider":"AntBio","version":"1.0","type":"link"}