| 摘要 |
FIELD: biotechnology. ^ SUBSTANCE: invention relates to cellular technology and can be used in medicine. Method for culturing human mesenchymal stem cells (hMSC) ex vivo involves at least two cycles for culturing nucleus-containing bone marrow cells in growth medium up to preparing the desired amount of mesenchymal stem cells. In the process of each cycle isolated cells are seeded in the corresponding cultural vessel and pH value of the growth medium is determined regularly, and the growth medium is changed at pH value less 6.0 or above 8.0 for the growth medium with pH value in the range 6.0-8.0, and cells not related to population of mesenchymal cells are removed regularly. The culturing process in each cycle is carried out up to formation of uniform monolayer of MSC population that is removed from cultural vessel by its treatment with 0.03-0.1% solution of trypsin. Before and after this treatment of MSC are washed out in isotonic solution from trace of the growth medium and trace of trypsin, respectively. Invention provides growing the effective therapeutic dose of MSC (at least 5 x 107 - 108) from 0.5-1.0 ml of a bone marrow puncture sample for rather short period (about 40 days). ^ EFFECT: improved culturing method of cells. ^ 9 cl, 5 ex |
