The invention belongs to the field of biological
medicine, and particularly discloses a culture method of domesticated mesenchymal stem cells (MSCs), which comprises the following steps: separating
plasma and
platelet lysis buffer from autologous
peripheral blood, and obtaining CTL and Treg through
magnetic bead sorting; the method comprises the following steps: sequentially co-culturing
umbilical cord MSCs for
three generations: co-culturing the P1 generation and CTL in a culture medium containing 30% of alpha MEM, 68% of DMEM / F12, 1% of
autologous plasma and 1% of
platelet lysis buffer, and adding 0.1-1 [mu] M of
resveratrol and 10-50 [mu] M of
spermidine; co-culturing P2 and Treg in a culture medium containing 50% of alpha MEM, 48% of DMEM / F12, 1% of
autologous plasma and 1% of
platelet lysis buffer; the P3 generation is cultured and harvested in 90% of alpha MEM, 5% of DMEM / F12 and 5% of PRP. The obtained MSCs
surface marker is CD105 < + > / CD90 < + > / CD73 < + > / CD45 <-> / CD34 <-> / HLA-DR <->, the expression quantity of CD24 is low (CD24low), the expression quantity of CD47 is high (CD47high), the expression quantity of PD-L1 is high (PD-L1 high), the IDO activity is larger than or equal to 25 U / mg, the
retention rate of the
lung after
vein transplantation is smaller than or equal to 12%, and the
survival period is larger than or equal to 28 days. The
cell is suitable for treating
ARDS or
hepatic fibrosis.