{"product_id":"breast-cancer-organoid-culture-medium-abs9526","title":"Human Breast Cancer Organoid Culture Medium","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\u003cstrong\u003e1 , Original generation: \u003c\/strong\u003e\u003cbr\u003e （ 1 The removed tissue was placed in a pre-cooled ( 2-8°C The tissue preservation solution was quickly transported to a clean laboratory for tissue processing and cell separation, photographed, and information recorded. \u003cbr\u003e （ 2 Prepare several petri dishes and add...  4℃ Precooled human breast cancer organoid tissue primary culture buffer ( abs9731 ) Backup. \u003cbr\u003e （ 3 The sample bottle is sterilized, the tissue is placed in a petri dish, washed three times with human breast cancer organoid tissue primary culture buffer, and then the tissue is cut into a volume of approximately [size missing] using ophthalmic scissors or a scalpel.  1-3mm\u003csup\u003e3\u003c\/sup\u003e Tissue blocks. \u003cbr\u003e （ 4 Human breast cancer primary tissue digestive fluid for tissue use ( abs9522 Digestion, 37℃ oscillation digestion 10-20min (Observe the digestion situation at any time during the digestion process). \u003cbr\u003e （ 5 Take a small amount of liquid and observe it under a microscope. A large number of individual cells or  70μm  After the following cell clusters, digestion was terminated by adding three times the volume of human breast cancer organoid tissue primary culture buffer. \u003cbr\u003e （ 6 ) Use 100μm The filtrate is filtered using a sieve with a certain pore size, and the filtrate is collected and then... 300g enrichment centrifugation 5min The supernatant was then removed, and human breast cancer organoid tissue primary culture buffer was added to resuspend and centrifuge. \u003cbr\u003e （ 7 Matrix rubber calculation: No. 6 After the step, observe the collected tissue volume and add 25 Matrix glue with twice the tissue volume ( abs9495 Resuspend the slab. \u003cbr\u003e （ 8 ） 24 Taking well cell culture plates as an example, each well is dispensed with gel. 25μL  Tissue matrix glue mixture for board laying ( 4℃ (Next operation). \u003cbr\u003e （ 9 Place the prepared culture plate in... 37℃ Incubator 10-15min Gelatinization was performed, and human breast cancer organoid culture medium was added (restored to room temperature) for culture. \u003cbr\u003e\u003cstrong\u003e2 Organoid subculture: \u003c\/strong\u003e\u003cbr\u003e （ 1 Remove the culture medium with a pipette and add to each well. 1-2mL 4℃ Organoid subculture buffer ( abs9730 ) Place 2min 。 \u003cbr\u003e （ 2 Gently blow the matrix glue with a pipette to collect... 15mL  In the centrifuge tube, 4℃ standing 10min 。 (each 6-8 (The holes are a group) \u003cbr\u003e （ 3 ） a : Insufficient number or small volume of organoids:   centrifuge 5min Discard the supernatant, add an appropriate amount of human breast cancer organoid subculture buffer, resuspend and transfer. 1.5mL  centrifuge tube, 300g centrifuge 5min Discard the liquid for the first 4 step. \u003cbr\u003e         b When the number or volume of organoids is large: Centrifuge 5min Discard the supernatant and add an appropriate amount of organoid passage digestive fluid ( abs9520 Digestion 2-3min Add human breast cancer organoid subculture buffer to terminate digestion and centrifuge. 5min Discard the mixture, add an appropriate amount of human breast cancer organoid subculture buffer, resuspend and transfer. 1.5mL  centrifuge tube, 300g centrifuge 5min Discard the liquid for the first 4 step. \u003cbr\u003e （ 4 After organoids were collected, matrix gel was added for resuspension, and each well was resuspended. 25μL  Matrix glue is laid on 24 In well cell culture plates, place in an incubator. 10-15min Add 500μL  Organoid medium for human breast cancer. \u003cbr\u003e\u003cstrong\u003e3 Organoid cryopreservation: \u003c\/strong\u003e\u003cbr\u003e （ 1 Remove the culture medium with a pipette and add to each well. 1-2mL 4℃ organoid subculture buffer placement 2min 。 \u003cbr\u003e （ 2 Gently blow the matrix glue with a pipette to collect... 15ml In the centrifuge tube, 4℃ standing 10min 。 (each 6-8 (The holes are a group) \u003cbr\u003e （ 3 Centrifugation 5min Discarding the supernatant, adding an appropriate amount of human breast cancer organoid subculture buffer for resuspension, 300g centrifuge 5min Discard the liquid. \u003cbr\u003e （ 4 Add an appropriate amount of organoid cryopreservation solution ( abs9519 ), gently blowing and resuspending, to 24 Taking well cell culture plates as an example: the density is 2 Individual hole cryopreservation 1 Tubes, volume per tube 1.4mL 。 \u003cbr\u003e （ 5 Mark the information, cool it down procedurally, and then transfer it to liquid nitrogen for long-term storage. \u003cbr\u003e\u003cstrong\u003e4 Organoid resuscitation: \u003c\/strong\u003e\u003cbr\u003e （ 1 ) take 10mL Human breast cancer organoid subculture buffer 15mL  In the centrifuge tube. \u003cbr\u003e （ 2 Remove the frozen organoid cells from the liquid nitrogen tank and quickly place them in...  37℃ Melt in a water bath. \u003cbr\u003e （ 3 During the water bath thawing process, the freezing tube needs to be gently shaken to ensure that the cryopreservation solution is...  1-2min It completely dissolves inside. \u003cbr\u003e （ 4 ) Rapidly transfer the lysed organoid cells to 15mL  Centrifuge tube, gently blow with a pipette 6-8 next time, 300g  centrifuge  5min The supernatant is then removed and the organoid cell pellet is collected. Add an appropriate amount of human breast cancer organoid subculture buffer to resuspend and transfer 1.5mL  centrifuge tube 300g centrifuge 5min 。 \u003cbr\u003e （ 5 Resuspend the matrix gel, per well 25μL  Matrix glue is laid on 24 In well cell culture plates, place in an incubator. 10-15min Gelation, addition 500μL  Organoid medium for human breast cancer. \u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003e\u003cstrong\u003eDescription\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd\u003e\u003cp\u003eThe human breast cancer organoid basic culture medium can be applied to the conventional culture and passage of human breast cancer-derived organoids, can maintain their growth vitality after passage, and does not need to add any other protein factors during the organoid culture process.\u003c\/p\u003e\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°C with a shelf life of 12 months.\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003c\/tbody\u003e\u003c\/table\u003e\u003c\/div\u003e","brand":"Absin","offers":[{"title":"100mL","offer_id":43214371618891,"sku":"abs9526-100mL","price":476.0,"currency_code":"USD","in_stock":true},{"title":"500mL","offer_id":43214371651659,"sku":"abs9526-500mL","price":1587.0,"currency_code":"USD","in_stock":true}],"url":"https:\/\/www.antbioinc.com\/products\/breast-cancer-organoid-culture-medium-abs9526","provider":"AntBio","version":"1.0","type":"link"}