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Description
Organotial kidney organ class training kitProduct Specification Description Composition: Component name Specification Human kidney cancer organoid medium A 100mL Primary culture buffer B 250mL Human kidney cancer primary tissue digestive juice C 30mL Organoid passage digestive juice D 30mL Tissue preservation solution E 100mL Organoid cryopreservation fluid F 20mL Organoid subculture buffer G 250mL Instructions 1, primary (1) tissue after sampling suggested 2 8 The tissues were stored under C
Product Specification
| Description |
Composition:
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| Instructions |
1, primary (1) tissue after sampling suggested 2-8° The tissues were stored under C conditions and quickly transferred to a clean laboratory for tissue processing and cell separation. The information was photographed and registered. (2) Several culture dishes were prepared and primary culture buffer B, precooled at 4 ° C, was added for later use. (3) The sampling bottle was disinfected, and the tissue was placed in the Petri dish. After washing three times with primary culture buffer B, the tissue was cut into tissue blocks of about 1-3mm3in volume using ophthalmic scissors or scalpel. (4) The tissues were digested with human renal cancer primary tissue digestion solution C and shaken at 37℃ for 15-25min (the digestion was observed at any time during the digestion process). (5) A small amount of liquid was observed under the microscope. When more single cells or cell clusters below 70um were observed under the microscope, triploid volume primary culture buffer B was added to terminate the digestion. (6) The filtrate was filtered by a sieve with a pore size of 100um. After collecting the filtrate, the supernatant was removed after enrichment and centrifugation at 300g for 5min. The supernatant was resuspended by adding primary culture buffer B, and the supernatant was removed after enrichment and centrifugation at 300g for 5min. (7) The cell precipitate contained red blood cells, the supernatant was discarded, 1-2 ml of red blood cell lysate was added for 1-2min, diluted to 10mL, and centrifuged at 300g for 5min. (8) Matrix glue calculation: The collected tissue volume was observed after step 6, and the paving plate was resuspended by adding matrix glue (abs9495) at 25 times the tissue volume. (9) For the example of 24-well cell culture plate, 25uL-30uL tissue matrix glue mixture was dispensed to each well for paving (Note: matrix glue was maintained at 0-4 ° C for operation). (10) The paved culture plate was placed in an incubator at 37 ° C for 10-15min. After the matrix gel solidified, 500-750uL of human renal cancer organoid medium A (returned to room temperature) was added for culture. 2, Organoid subculture (1) The medium was aspirated by pipetting gun, and 1-2mL of 4 ° C organoid subculture buffer G was added to each well for 2min. (2) The matrix glue was gently blown by a pipetting gun, collected in a 15mL centrifuge tube, and allowed to rest for 10-15min at 4 ° C. (every 6-8 Wells are a group) (3) a: When the number of organoids is insufficient or the volume is small: Discard the supernatant by centrifugation at 300g for 5min, add 1mL organoid passage culture buffer G, resuspension and transfer into 1.5mL centrifuge tube, and discard the liquid by centrifugation at 300g for 5min for the fourth step. b: When organoids are large in number or size: Centrifugation at 300g for 5min to discard the supernatant, add 1-2mL organoid passage digestion solution D for 2-4min, add 3 times the volume of organoid passage culture buffer G to terminate digestion, centrifugation at 300g for 5min to discard the supernatant, add an appropriate amount of organoid passage culture buffer G to resuspend and transfer to a 1.5mL centrifuge tube. The supernatant was discarded by centrifugation at 300g for 5min, and the fourth step was carried out. (4) After organoids were collected, they were resuspended by adding matrigel, and 25uL of Matrigel per well was spread in 24-well cell culture plates and placed in an incubator for 10-15min. After the Matrigel had set, 500 to 750uL of human renal cancer organoid medium A was added to each well. 3, organoids were frozen (1) The medium was removed by suction with a pipetting gun, and 1-2 ml of 4 ° C organoid passage culture buffer G was added to each well and placed for 2min. (2) The matrix glue was gently blown by a pipetting gun, collected in a 15mL centrifuge tube and allowed to rest for 10min at 4 ° C. (every 6-8 Wells were a group) (3) The supernatant was discarded by centrifugation for 5min, then an appropriate amount of organoid passage culture buffer G was added and resuspended again, and the liquid was discarded by centrifugation at 300g for 5min. (4) Add 2mL organoid cryopreservation solution F to every 3 Wells, gently blow and mix, and transfer to cell cryopreservation tubes, 1mL per tube. (5) Make the labeling information, after the program cooling, move to -80° C refrigerator, 48h later, transferred to liquid nitrogen for long-term storage. Or put it into the 4℃ refrigerator for 40min, put it into the -20℃ refrigerator for 2h, move it to the -80℃ refrigerator, and put it into the liquid nitrogen tank for storage after 48h. 4, organoid resuscitation (1) 10mL organoid subculture buffer G was placed in a 15mL centrifuge tube. (2) The frozen organoid cells were removed from the liquid nitrogen tank and quickly thawed in a 37℃ water bath. (3) During the water bath melting process, it is necessary to gently shake the freezing tube to ensure that the frozen solution is completely melted in a short time. (4) The dissolved organoid cells were quickly transferred to a 15mL centrifuge tube, gently blown with a pipette for 6-8 times, centrifuged at 300g for 5min, and then the supernatant was removed and the organoid cell precipitate was collected. An appropriate amount of organoid passage culture buffer G was added and resuspended, then transferred to a 1.5mL centrifuge tube and centrifuged at 300g for 5min, and the supernatant was discarded. (5) Add 100ul of Matrigel to each tube of cryopreserved tube and resuspend, 25uL of Matrigel in each well was spread in 24-well cell culture plate, placed in the incubator for 10-15min to gel, and 500-750uL of human renal cancer organoid medium A was added. |
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| Storage Temp. | Stored at 4 ° C, valid for 3 months; Stored at -20 ° C, valid for 1 year, see label for details. |
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4.5 ★★★★★
Based on 9 reviews
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Product Reviews
★★★★★ 4
Good!!!
Size: Medium, Color: Avocado
Excellent quality and durability.My pets really enjoy it and it's almost impossible to damage it!!
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on May 31, 2026
★★★★★ 5
By far the best chew toy for aggressive dog chewers
Style: Dog Stick-Blue
Definitely high quality very durable very indestructible chew toy my dog destroys just about every toy I get him and this I will say he’s haven a hard time with destroying this 1 so that being said all I can say is highly recommend this chew toy for your aggressive dog chewer
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Reviewed in the United States on June 4, 2026
★★★★★ 5
Lifesaver for playtime
This toy is AMAZING. My dog still loves to play with it every week, and somehow by magic it hasn't been broken once. We have had it for 9 months, that is a record for toys in our house. If you are on the fence, just get it already. Ours doesn't squeak anymore obviously, but we still play tug a war with it a lot and it flops and bends and bounces just fine!!
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Reviewed in the United States on April 16, 2026
★★★★★ 5
Tough and lasts
Bought this toy Sept 2025. This has been my dogs favorite toy. It was hard when we bought it and he didn’t like it at first. Once he started playing with it, it became the toy of choice. It has softened a lot and loves it even more. First thing we greeted with in the morning is Dude with the yellow toy. I’m going to buy another to have if I think it gets too soft.
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Reviewed in the United States on April 17, 2026
★★★★★ 4
He loved it
Is it durable? Not at all. Did he love it? While it lasted.
Our dog is a monster chewer with toys so I go into every purchase knowing we might get 8 hours out of a toy if we're lucky, depending on visible durability. We don't even buy stuffies anymore because it's just not smart. He loved the squeak while it lasted, it's large so we never have to worry about him choking on it while running/being dumb. If you have a dog that will eat pieces of things as they destroy toys, this is not the toy for them. Ours plucks it apart piece by piece, but leaves the bits near one of us (LUCKILY!). If he didn't, we'd never buy this again. The missing star is just for lack of durability. Even for a rubber toy it felt like it didn't last very long.
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Reviewed in the United States on December 12, 2025