The invention discloses an umbilical cord mesenchymal stem cell (MSC) separation culture and amplification method. The umbilical cord MSC separation culture and amplification method comprises the steps of (1) clearing, (2) treatment, (3) culture, and (4) inoculation and passage. The umbilical cord MSC method obtained by adopting a tissue mass separation method is simple, the cost is low, cell activity is good, purity is high, the final harvested cell concentration is high, the obtained MSC method is simple, MSCs can be quickly and efficiently separated, the culture time is shortened, after culture is conducted for 4-5 days, it can be seen that the cells climb out from tissue masses, the final harvested cell concentration is high, and the entire umbilical cord MSC culture process can be standardized, programmed and normalized.