The invention relates to a high efficiency fetal calf serum substitute. The high efficiency fetal calf serum substitute is composed of recombinant human albumin, recombinant human transferrin, galactose, lipid, a trace element compound and vitamins, a weight ratio of the recombinant human albumin to the recombinant human transferrin to the galactose is 2:50:(15-25), and the lipid is a cholesterol, linoleic acid and linolenic acid mixture. The fetal calf serum substitute provided by the invention avoids infectious disease propagation risk and foreign protein induced allergy, increases the in vitro culture survival rate and the increment multiple of human lymphocytes to 96% and 28 respectively, improves the supply of the lymphocytes, reduces the culture cost, and facilitates large scale use.