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Description
Human Liver Organoid Culture Medium KitProduct Specification Usage 1. Primary (1) the tissue after sampling must be placed in the pre cooled (2 8 c) sampling bottle of tissue preservation solution e, and quickly transferred to the clean laboratory for tissue processing and cell separation, taking photos and registering information (2) prepare several Petri dishes and add 4 pre cooled primary culture buffer B for standby (3) disinfect the sampling bottle, put the tissue into the Petri dish,
Product Specification
| Usage |
1. Primary (1) the tissue after sampling must be placed in the pre cooled (2-8° c) sampling bottle of tissue preservation solution e, and quickly transferred to the clean laboratory for tissue processing and cell separation, taking photos and registering information (2) prepare several Petri dishes and add 4 ℃ pre cooled primary culture buffer B for standby (3) disinfect the sampling bottle, put the tissue into the Petri dish, wash it three times with primary culture buffer B, remove impurities, and cut the tissue into a tissue block with a volume of about 1-3mm3by using ophthalmic scissors or scalpel (4) the tissue is digested with human normal liver primary tissue digestive solution C, and then shaken for 10-20min at 37 ℃ (observe the digestion at any time during the digestion process) (5) take a small amount of liquid and observe it under the microscope. After observing more single cells or cell clusters below 70UM under the microscope, add three times the volume of primary culture buffer B to stop digestion (6) filter with a 100um pore size screen, collect the filtrate, remove the supernatant after centrifugation at 300g for 5min, add primary culture buffer B and resuspend for centrifugation (7) Matrigel calculation: after step 6, observe the collected tissue volume, add 25 times the tissue volume of Matrigel (abs9495) to resuspend the plate (8) take a 24 well cell culture plate as an example, dispense 25ul of tissue Matrigel mixture per well for plating (operate at 4 ℃) (9) put the laid culture plate into a 37 ℃ incubator for 10-15min to form a gel, and add human normal liver organoid medium a (restore room temperature) for culture 2. Organoid subculture (1) pipette the culture medium, add 1-2ml of 4 ℃ organoid subculture buffer g to each well and place it for 2min (2) pipette the Matrigel gently, collect it in a 15ml centrifuge tube, and let it stand at 4 ℃ for 10min. (each 6-8 well is a group) (3) a: the number of organoids is insufficient or the volume is small: centrifuge for 5min to discard the supernatant, add an appropriate amount of organoid subculture buffer g to resuspend and transfer into a 1.5ml centrifuge tube, centrifuge for 5min at 300g to discard the liquid for step 4  &Nbsp&Nbsp&Nbsp b: When the number of organoids is large or the volume is large: centrifuge for 5min to discard the supernatant, add an appropriate amount of organoid passage digest D for digestion for 2-3min, add organoid passage culture buffer g to stop digestion, centrifuge for 5min to discard the mixture, add an appropriate amount of organoid passage culture buffer g to resuspend and transfer into a 1.5ml centrifuge tube, centrifuge for 5min at 300g to discard the liquid for step 4 (4) after organoids are collected, add Matrigel to resuspend, lay 25ul Matrigel per well in a 24 well cell culture plate, place in the incubator for 10-15min, and add 500ul human normal liver organoid culture medium a 3. Organoid cryopreservation (1) pipette the culture medium, add 1-2ml of 4 ℃ organoid subculture buffer g to each well and place it for 2min (2) pipette the Matrigel gently, collect it in a 15ml centrifuge tube, and let it stand at 4 ℃ for 10min. (each 6-8 well is a group) (3) centrifuge for 5min to discard the supernatant, add an appropriate amount of organoid subculture buffer g to resuspend, and centrifuge for 5min at 300g to discard the liquid (4) add an appropriate amount of organoid cryopreservation solution F, gently blow and resuspend. Take a 24 well cell culture plate as an example: the density is 2 wells and 1 tube is frozen, with a volume of 1.4ml per tube (5) mark the information, and after cooling down the program, transfer it to liquid nitrogen for long-term storage 4. Organoid resuscitation (1) take 10ml of organoid subculture buffer g and put it into a 15ml centrifuge tube (2) take out the frozen organoid cells from the liquid nitrogen tank and quickly put them into a 37 ℃ water bath pot for melting (3) during the water bath thawing process, it is necessary to gently shake the freezing tube to ensure that the frozen solution is completely thawed in 1-2min (4) quickly transfer the dissolved organoid cells to a 15ml centrifuge tube, gently blow 6-8 times with a pipette, centrifuge at 300g for 5min, and then remove the supernatant and collect the organoid cell pellet. Add an appropriate amount of organoid subculture buffer g to resuspend and transfer into a 1.5ml centrifuge tube. Centrifuge for 5min at 300g (5) resuspend Matrigel, lay 25ul Matrigel per well in a 24 well cell culture plate, place in an incubator for 10-15min to form a gel, and add 500ul human normal liver organoid culture medium a. |
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| Description | Human normal liver organoid culture medium is a medium that promotes the formation and expansion of human normal liver organoids in vitro. The product is a sterile liquid mixing system, containing amino acids, vitamins, organic and inorganic compounds and growth factors necessary to maintain the growth of target cells. The unique formula of this medium can provide a suitable nutritional environment for cells and promote the growth and expansion of human normal liver organoids in vitro Composition:
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| Storage Temp. | 4° C. 3months; -20° C. 1 year, see label for details |
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4.2 ★★★★★
Based on 11 reviews
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Product Reviews
★★★★★ 1
Would not buy again
My dog loves chew toys and was very excited when I opened the package. She played with it for about 10 minutes and now she wont touch it. It does seem very durable as she chewed on it and there was little damage.
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Reviewed in the United States on May 31, 2025
★★★★★ 3
Long lasting, not super appealing to my dogs.
Objective Data: I have two 50 pound dogs; a 10mos goldendoodle and a 7yo viszla. The puppy is a moderate chewer, the senior dog takes chewing in binges. He will occasionally get something and work it over for awhile and then not seek out anything to chew for a couple of days.
Initially these went over well, but neither of my dogs pursue these much after a few weeks. The puppy will occasionally bring one over to briefly chew laying at my feet.
Even with the initial, enthusiastic chewing of "new" toys these are still very intact!! These are going to last such a long time.
**CAUTION** if you have beige carpet I don't recommend these toys for your home. They blend in to the color of our carpet and I cannot tell you how many times I have rolled my ankle or stubbed my toe kicking one across a dim/dark room. I would rather walk barefoot through a minefield of legos.
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Reviewed in the United States on March 21, 2025
★★★★★ 5
odd shape but dogs are constantly chewing on it
seems to be an odd shape but my large dogs constantly chew on this. holds up to chewing well.
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Reviewed in the United States on October 17, 2025
★★★★★ 4
Keeps my dog busy
My dog has so much fun getting peanut butter out of this chew! It has small holes so it makes them put a little more effort than a typical chew. I would say it keeps them occupied for at least 15-20 minutes and they get pretty tired after. I think it's a good size for my medium dog. One thing i wish they did was making it with a silicone rather than nylon. It does get chewed off and I don't like the fact that they are ingesting them. All in all it's a great design to keep than busy.
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Reviewed in the United States on March 17, 2025
★★★★★ 5
Seriously durable
I have 3 XXL dogs (how convenient this comes in a three pack!). They will destroy many toys marked for aggressive chewers, but not these. Plus, they are good sized, leaving room for me to hold them while they are chewing when I elect to do that.
I don't really smell anything in the toy itself. Maybe the dogs do, but not enough to keep their interest more than a few minutes. However, when I smeared it with peanut butter, it was a whole new story. All 3 of them LOVED that and spent some time getting every last morsel of peanut butter out of the toy. In the meantime, because of the honeycomb texture, I feel they were getting a decent dose of tooth cleaning as well.
I wish the texture were a little deeper to fit more peanut butter, but I suppose it is probable that would make the surface texture less durable.
My dogs like them, so I do too!
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Reviewed in the United States on February 20, 2025
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