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Description
Human Lung Cancer 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 human lung cancer primary tissue digestive solution C, and the shaking digestion was carried out 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 human lung cancer 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 lung cancer 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 the Matrigel, lay 25ul of Matrigel per well in a 24 well cell culture plate, place it in an incubator for 10-15min to form a gel, and add 500ul of human lung cancer 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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4.6 ★★★★★
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Product Reviews
★★★★★ 5
Best Balls Ever!!!
Size: Medium
These are the best balls ever! We have 2 very active ball loving dogs and these bounce very well and they actually hold up with active play and chewing. They still look brand new after daily use for months!
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Reviewed in the United States on May 15, 2026
★★★★★ 5
Finally a ball my power chewers cannot destroy
Size: Medium
I have two dogs, an American Bully and an American Pit Bull Terrier, and both of them love to chew. Most balls last a very short time around here before they are shredded or missing big chunks. This one has actually held up. They chase it, tug on it and sit and chew on it, and it is still in one piece with no pieces breaking off.
One of my favorite things is being able to put food inside. I add some kibble or small snacks, and they will happily sit and work on it for a long time. It keeps them busy, gives them something safe to chew, and they get a little reward while they play. When they are done, I just rinse it out and it is ready to go again.
If you have strong jawed dogs that usually destroy toys, this ball is worth trying. Mine really enjoy it and I feel better knowing they finally have a ball that can keep up with them.
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Reviewed in the United States on November 22, 2025
★★★★★ 5
My dog LOVES this ball
Size: Medium, Size: Medium
Does your dog destroy tennis balls in seconds?
Yes? Then your dog NEEDS this ball!
It's squishy like a tennis ball, fits in ball launchers (even the cheap ones) and bounces when it hits the ground.
My dog has been literally playing with this ball all day and not a single mark on it!
The only bad thing is that it does not float.
Definitely worth the money!
Put it in your cart!
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Reviewed in the United States on March 8, 2026
★★★★★ 5
Perfect for ball demolishers
Size: Medium
The dogs love the chuck it brand. I don’t know what it’s made of but they go crazy over them. Will not play with another ball. Wonder if it’s “laced” with an addictive scent. Have pits. Very durable. Not able to chew to pieces. Last for about 6 months before there is a small break. I keep a back up because when the ball goes missing the house is in chaos. 😆 🤪
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Reviewed in the United States on March 27, 2026
★★★★★ 5
Most durable ball!
Size: Medium
This are my dogs fav ball 😍 they last forever!
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Reviewed in the United States on April 23, 2026
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