Umbilical cord mesenchymal stem cells(hUC MSCs) are a type of stem cell with multipotent differentiation ability extracted from the umbilical cord of newborns. Umbilical cord derived mesenchymal stem cells not only maintain the biological characteristics of mesenchymal stem cells (surface antigen markers, with the ability to differentiate into osteoblasts, chondrocytes, and adipocytes), but are also safer and more primitive compared to other sources of mesenchymal stem cells, with stronger proliferation, differentiation, and immune regulatory abilities.
In addition, umbilical cord mesenchymal stem cells have the characteristics of convenient sampling and easy isolation, low immunogenicity, no ethical restrictions, a large number of available cells, and the ability to achieve large-scale standardized production, which have broad clinical application prospects. This product has undergone strict quality control and testing to ensure its safety and effectiveness.
(1) When the cell density reaches 80% -90%, passage can be carried out (note that the cells cannot be too dense, otherwise they are prone to differentiation). Discard the culture supernatant and rinse the cells 1-2 times with DPBS that does not contain calcium or magnesium ions;
(2) Add 1mL of recombinant trypsin to the culture bottle to allow the digestion solution to infiltrate all cells. Place the cells in a 37 ℃ incubator for digestion for 1-3 minutes and observe under a microscope until most of the cells become round and shed (depending on the condition of the cells).
(3) Add and transfer to sterile centrifuge tubes, centrifuge at 1000rpm for 5 minutes, discard 3ml of MSC medium to terminate digestion. Gently mix and remove the supernatant, then add 1-2ml of MSC medium and blow evenly.
(4) Passage should be carried out in a ratio of 1:3 or 1:4, or cells should be counted and seeded into new culture bottles at a concentration of 5-8 × 103 cells/cm2. After supplementing with the corresponding volume of MSC medium, the cells should be cultured in a 37 ℃, 5% CO2 incubator.
(5) Change the medium every 2-3 days and culture for 2-4 days until the cell growth density reaches around 80% before proceeding with subsequent passaging or cryopreservation operations.
Digestion time: Pancreatic enzyme digestion should be controlled for about 1 minute to avoid excessive digestion and cell damage.
Inoculation density: Cultivate in a ratio of 1:3 to 1:4 according to the base area to avoid over density or over dilution.
Generation selection: It is recommended to use low generation cells (such as P2 generation) to ensure experimental effectiveness.
Specialized culture medium: Use specialized culture medium containing growth factors to promote cell proliferation and differentiation.