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Description
Human Endometrial carcinoma 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 with primary culture buffer B for three times, and then cut the tissue into a tissue block with a volume of about 1-3mm3using ophthalmic scissors or scalpel (4) the tissue was digested with human endometrial cancer 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 endometrial 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 endometrial 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 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 human endometrial 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
★★★★★ 1
DO NOT BUY (lasted less than 7 months)
Size: 11, Color: Black, Size: 11, Color: Black
I bought these for work as a bartender in August of 2025. By December, the tops were scuffed, frayed, and peeling beyond repair, and by February, the soles were coming off. I’ve never had a pair of shoes with such low quality.
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Reviewed in the United States on April 11, 2026
★★★★★ 5
Most comfortable shoes, right out of the box!
Size: 8.5, Color: Cognac Napa Leather
Oh my! So, my ordering of shoes online has been hit or miss - 1/2 size too big or too small; too wide or not wide enough. Enter these Marc Joseph shoes… such a game changer. They fit like a glove, exceptionally comfortable, and easy to slip in and out of. I like them so much that I am shopping for another pair. So far, the size, quality, fit, feel, look, and comfort make this pair my dedicated first pick. Btw, I have been wearing them for six hours and they are still very comfortable! I highly recommend these shoes.
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Reviewed in the United States on September 28, 2025
★★★★★ 5
Me gustaron
Size: 9.5 Wide, Color: Black
Llego exactamente la talla que pedí, son como se ven en la foto!
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Reviewed in the United States on June 3, 2026
★★★★★ 5
Comfortable dress shoe.
Size: 7, Color: Brown Napa Leather
Finally a comfortable dress shoe. It's both comfortable to walk in and looks and wears like a more traditional dress shoes for the office.
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Reviewed in the United States on May 6, 2026
★★★★★ 5
Slip-on ready and great quality
Size: 9, Color: Black
These shoes were stuff for the first half hour but then feel great. Well worth the chance and money. Great quality and slip-on easy
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Reviewed in the United States on May 22, 2026