The invention discloses a serum-free culture scheme for human umbilical cord mesenchymal stem cells (hUC-MSC). According to the scheme, the hUC-MSC are cultured by adopting a stepped method, wherein the hUC-MSC are cultured with a TME culture medium for 3-4 hours to promote adherence of the hUC-MSC, and then the hUC-MSC are cultured in a TMD culture medium for rapid amplification. According to the TME-TMD serum-free stepped culture method, the problems that in a serum-free culture method for the hUC-MSC, the cell adherence capacity is poor, multiplication is slow and differentiation is prone to occur are better solved, and a base is laid for a long-term stable serum-free culture technique for the hUC-MSC.