Objective To select a safe,low-cost basal medium in combination with platelet lysate(PL),suitable for the growth of human umbilical cord mesenchymal stem cells(hUC-MSCs)with no influence on the functions of cells during the cultivation process,and to use it for the culture of a large number of high-generation hUC-MSCs.Methods Three different basal media,MSCBM,α-MEM and IMDM,were mixed with PL UltraGROG Advanced respectively to culture hUC-MSCs from P0 to P8.The cell morphology,cell expansion quantity,proliferation function,differentiation function and cell surface markers of different generations were compared to determine the optimal basal medium suitable for hUC-MSCs cultivation.Results The morphology of P1-P5 cells cultured in the three media was uniform and radial,with no significant difference between them and the standard cells(tiMDMVSα-MEM=0.159,tMSCBMVSα-MEM=0.147,tIMDMVSMSCBM=0.161;each P＞0.05).The morphology of P6-P8 cells cultured in MSCBM and α-MEM showed no significant difference compared with that of P0-P5 cells(tIMDMVSMSCBM=0.132,tIMDMVSα-MEM=0.128;each P＞0.05).The positive expression rate of CD73,CD90 and CD105 all reached about 99％in P1-P8 cells cultured in MSCBM,α-MEM and IMDM,while the expression of CD34 and CD45 was negative with the expression rate of lower than 2％,and there was no significant difference among different media(tIMDMVSα-MEM=0.102,tMSCBMVSα-MEM=0.106,t1MDMVSMSCBM=0.113;each P＞0.05).The clonal formation ofP8 cells cultured in the three media was different,the numbers of single clonal cluster and MSCBM cells in clonal cluster were higher than those of the same generation cells cultured in α-MEM and IMDM(t=0.023 and 0.049,respectively,each P＜0.05).The proliferative function of cells cultured in the three media from high to low was MSCBM,α-MEM and IMDM,among which,the proliferation function of cells cultured in MSCBM was significantly higher than that in IMDM(t=0.041,P＜0.05),while there was no significant difference between MSCBM and α-MEM(t=0.211,P＞0.05).The result of doubling time(DT)was consistent with the proliferation function.The cells of the same generation cultured in the three media all had strong differentiation function,and there was no significant difference in the number of osteoblasts and chondroblasts differ-entiated from P8 cells(tIMDM VSo-MEM=0.119,tMSCBM VSα-MEM=0.112,tIMDM VS MSCBM=0.111,each P＞0.05).The number of adipoblasts differentiated from P8 cells cultured in MSCBM was higher than that from P8 cells cultured in α-MEM(t=0.036,P＜0.05),and the number of adipoblasts differentiated from P8 cells cultured in α-MEM was higher than that in IMDM(t=0.031,P＜0.05).Conclusion In this study,the optimal culture system of hUC-MSCs was MSMBM with 5％UltraGROG Advanced,followed by α-MEM with 5％UltraGROG Advanced,and these two serum-free culture systems were suggested to be selected for large-scale culture of MSCs.
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