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Description
Mouse 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 was digested with the digestive solution c of primary mouse normal liver tissue, and then it was shaken at 37 ℃ for 10-20min (the digestion condition was observed 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 mouse 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 were 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 of mouse 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 of mouse normal liver organoid culture medium A. |
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| Description |
Composition:
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| Storage Temp. | 4° C. 3months; -20° C. 1 year, see label for details |
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4.5 ★★★★★
Based on 14 reviews
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Product Reviews
★★★★★ 5
Indestructible ball….Destroyed my dog’s goals
Color: Brown
I have a golden retriever that would rather watch the world burn with every toy I’ve ever gotten him than play nice with any of them.
He burns through toys faster than a cardboard bonfire.
However, the trap was set…I bought this as a last ditch effort to show that four legged champion of destruction that he could be bested…
I remember it to this day..
It was a Tuesday - raining
The ball came in the mail and as I nervously unwrapped and opened it, it was as if he knew a new challenger stepped in the ring as he came barreling towards the front door where I stood.
Not today I told myself as I stood with confidence with this ball of justice in front of me.
The usual sit and stay kept him at bay until I tossed it to him in which he had turned into a frantic honey badger shaking the ball and chewing ferociously he wanted the squeaking thing inside that doesn’t squeak but sounds like an ugly cry.
However, it is next Tuesday and the ball stands victorious
My dog has been bested and now understands the chain of command.
He respects the ball but will always come back for more until he feels that he has won
All in all absolutely buy this for aggressive chews and toy playing dogs.
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Reviewed in the United States on March 26, 2024
★★★★★ 5
Broke the first day
I purchased the owl ball because of other dog chew toys I purchased from Amazon. Those other chew toys are still holding together. The main reason I got the owl ball is due to the different noises it makes other than the regular squeak noise. After giving the toy to my dog he loved it right away. He especially liked the noise it made. After a few hours, I noticed he was chewing a white tube-like device. I took this away from him and determined it was the noise maker in the owl ball. I examined the owl ball and noticed the glue used to keep the ball cap shut did not hold. On closer inspection, I noticed what looked like rubber cement was used to hold the cap on but if it was rubber cement it was used improperly. Anyway, I just reglued the cap correctly and now it's just a ball because my dog destroyed the noise maker.
OK, Since I posted this review I was contacted by the seller. They offered to give me a full refund for the owl ball. I accepted the offer. So, I will try it again, but buy a different color owl this time. I never expected a refund since it's just a ball now, my dog still plays with it and it is holding up very well. The material it is made from is pretty sturdy.
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Reviewed in the United States on September 4, 2024
★★★★★ 3
Squeaker not working!
Color: Yellow
I'll give it 3 stars because I love the design and it's sturdy and great for dogs to hold/launch. That said, the squeaker doesn't really work... I tried everything but it just doesn't make any sound. My dog wasn't really interested but when it did make a slight sound he approached it. Once it stopped he lost interest. But he's very difficult to please😆 so if you have a normal dog, he'll probably like it!
I hope the manufacturer fixes the squeaker issue because apparently many people are having the same issue, which is a shame because the product is clearly well made!
I'm on the fence about returning it just because the quality it's amazing(squeaker issue aside)!
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Reviewed in the United States on September 19, 2025
★★★★★ 4
Sturdy rubber ball. Noise maker could be improved
Color: Red
I would have given it five stars because of how much, my dog loved it in the beginning.
And shut us up holds true as a sturdy rubber ball. My only complaint is at with one of my dogs being a heavy chewer, she was able to bust the noise making part. The ball never broke open or anything. It just no longer makes noise. You can hear the piece rattling on the inside. The ball itself is still solid tho
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Reviewed in the United States on March 6, 2026
★★★★★ 1
POOR QUALITY WHILE OVER PRICED
Color: Brown
Like so many others said, the sound box in this ball is faulty. lasted ONLY for 30seconds. My dog loves the giggle ball I purchased prior so I knew she would like this as well. Well, she did, for that whole 30 seconds. Both of us were so disappointed we could not further our play. Plus this ball was so much smaller than expected for the cost. I would not have minded the cost had this ball been on the ball! Don't purchase this item. Buy the giggle ball instead if you haven't already.
UPDATE...
Ran across this message at a later date and I would like to thank the sellers at Mewlmart for their response and showing the best business practices as stated in their response below.
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We sincerely apologize for any inconvenience regarding your order and would like to offer you a full refund for your purchase.
If you would like a full refund: please reply 'Full refund'.
We hope that you give us another chance. Thank you very much.
Best regards,
Mewlmart
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Only thing is I wish I had seen their message earlier, as I believe the window of opportunity for a refund may no longer be applicable. But who knows, they may reach out and still honor it when they review this review. Will update if that happens as it is worth mentioning the extra mile they may take. I am sure I would just then turn around and buy another product from them, giving them another try:)
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Reviewed in the United States on January 2, 2024