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Description
Organotial Human Normal esophageal organoid Culture KitProduct Specification Description Composition: Component name Specification Human esophagus cancer organoid medium A 100mL Primary culture buffer B 250mL Human esophagus 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) The harvested tissues should be placed in precooled (2 8 C)
Product Specification
| Description |
Composition:
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| Instructions |
1, primary (1) The harvested tissues should be placed in precooled (2-8° C) In the sampling bottle of tissue preservation solution E, the tissue was quickly transferred to a clean laboratory for tissue processing and cell separation, photographed and recorded. (2) Prepare several culture dishes and add primary culture buffer B precooled at 4 ° C 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 normal human esophageal primary tissue digestion solution C and shaken at 37℃ for 20-25min (the digestion was observed at any time during the digestion). (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, and the filtrate was collected. After enrichment and centrifugation at 300g for 5min, the supernatant was removed, and the primary culture buffer B was added and resuspended for centrifugation. (7) Matrix glue calculation: The volume of collected tissue was observed after step 6, and matrix glue (abs9495) was added 25 times the tissue volume to resuspend the paving plate. (8) For the example of 24-well cell culture plates, 25uL of tissue matrix glue mixture was dispensed to each well for paving (operation at 4 ° C). (9) The paved culture plate was placed in an incubator at 37 ° C for 10-15min to become gelation, and human normal esophageal organoid medium A was added for culture (return to room temperature). 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 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) a: When the number of organoids was insufficient or the volume was small: the supernatant was discarded by centrifugation for 5min, then an appropriate amount of organoid passage culture buffer G was added and resuspended into a 1.5mL centrifuge tube, and the liquid was discarded by centrifugation at 300g for 5min for the fourth step. b: When organoids are large in number or size: The supernatant was discarded by centrifugation for 5min, then the organoid passage digestion solution D was added for 2-3min, then the organoid passage culture buffer G was added to terminate the digestion, the mixture was discarded by centrifugation for 5min, then the organoid passage culture buffer G was added and the mixture was resuspended and transferred to a 1.5mL centrifuge tube, and the liquid was discarded by centrifugation at 300g for 5min for step 4. (4) After the organoid was collected, 25uL of Matrigel was added to the 24-well cell culture plate, and 500uL of human normal esophageal organoid medium A was added to the incubator for 10-15min. 3. Organoids were frozen (1) The medium was removed by suction with a pipetting gun, and 1-2 ml of 4℃ 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 appropriate amount of organoid cryopreservation solution F, gently blow and resuspend, take 24-well cell culture plate as an example: the density is 2 Wells and freeze 1 tube, each tube volume is 1.4mL. (5) After making the labeling information and cooling, the samples were transferred to liquid nitrogen for long-term storage. 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 within 1-2min. (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 into a 1.5mL centrifuge tube and centrifuged at 300g for 5min. (5) Matrigel was resuspended, 25uL of Matrigel per well was spread in A 24-well cell culture plate, placed in an incubator for 10-15min to gel, and 500uL of human normal esophageal 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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4.1 ★★★★★
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Product Reviews
★★★★★ 5
Really comfortable!!!
Size: 11, Color: New Black05
These tennis shoes are so comfortable!!!
I would recommend anybody to get these, especially if you do a lot of walking they’re very light but have a good heal support and the soul is nice and soft and cushy..
They’re also so cute. I got the blue ones and red also.
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Reviewed in the United States on April 6, 2026
★★★★★ 5
i would recommend these shoes
Size: 11, Color: Black White 05
great shoes for the price. very comfortable...
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Reviewed in the United States on May 14, 2026
★★★★★ 5
Great Fit, Lightweight, and Totally Quiet
Size: 11, Color: New Black05
Great fit and very comfortable once you pick the right size. Lightweight, breathable, and completely silent on hard floors. No squeaks or “suction” sounds like some reviews claimed. Normal packaging, normal opening, no issues at all. Excellent value for the price.
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Reviewed in the United States on May 18, 2026
★★★★★ 5
Most comfortable shoes I've ever worn
Size: 11, Color: New Black05
First off, I was not expecting much buying a no-name shoe online, especially for the price. Boy was I in for a surprise...
I've owned these for almost a full year, and can confidently say these are the most comfortable shoes I've ever worn. I normally wear a size 11 and these fit me perfectly. Prior to this the most comfortable shoes I had were some Cole Haans with the Nike air technology. These shoes are so lightweight, so comfortable, I can wear them all day long without feeling a thing. Whenever I (rarely) need to wear a different pair of shoes, I am immediately reminded of how much I used to hate wearing shoes and couldn't wait to take them off.
Based on the price, I expected them to wear down quickly. However they are still going strong. Running, hiking, long walks, biking, worn to office / casual events. These shoes have been soaked in mud multiple times, but clean up easily by just throwing them in the washing machine and then the dryer. The soles and the insoles have both worn down a bit over the past year; at this rate I expect to get 1.5 to 2 years out of them before needing to be replaced. So far I must have put at least a thousand miles on these shoes based on how much I wear them.
They look good too. I have the all-black pair, and they look decent with jeans and a button up shirt for a night out, but also look natural in an athletic setting with shorts and no socks. Very versatile and subtle.
Now for the cons, which are minor. The pattern on the bottom of the sole is prone to some squeaking/suction sounds at first. However after about a month some of the ridges wore down just enough that it was no longer a problem. Second: they are not as breathable as I would have liked. For example, when running or biking, you don't feel any air on your feet. Wearing them out in the sun on a hot summer day, your feet will get a bit stuffy. In fact the fabric is woven tight enough that I wear them in the winter and in the rain, and don't get too cold or wet. So I do wish they made a more breathable version for summer time use.
All that being said, these are still the best pair of shoes I've ever had. Ultra light weight, comfortable, and can be worn all day long without feeling any fatigue whatsoever.
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UPDATE: It's been 1 year and 9 months, and the soles on these shoes are fully worn out and ready for replacement. Considering I've put between 1,000-2,000 miles of walking in these, that's pretty good. If anyone from Feethit is reading this, here are my top suggestions to perfect this shoe:
1) Easy fix - change the tread pattern. The current pattern causes suction cup effect on hard floors until it starts to wear down. Simply add some gaps in the circle pattern in the tread and it will remedy that issue.
2) Most importantly, put a thin piece of durable rubber on the bottom of the sole. The sole does wear down after some time and lose traction, especially on smooth wet floors (e.g. in the rain, in a kitchen, etc.). A thin piece of durable rubber would add more traction and also extend the life of the cushion-like sole (which is fantastic, don't change it!). This would easily be worth an extra $5-$10 in the price.
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Reviewed in the United States on August 4, 2023
★★★★★ 4
Nice shoes, despite the squeak and inconsistent sizing
Size: 12, Color: Black White 05
Excellent shoes at this price point. Quite comfortable. They do tend to have a nasty squeak on tile floors, sort of like a suction cup being engaged and disengaged.
I bought the navy blue and went back and bought the black. I have to say that the size varied greatly. With the second pair, I have to be sure to wear thin socks and even then, I struggle to get them on, despite having an extremely narrow foot. I'm not sure that a person with an average width foot could wear them. Buyer beware. You may need to go up a size.
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Reviewed in the United States on February 13, 2026