三种培养基对人脐带间充质干细胞培养生长和数量影响的比较

Comparison on the effects of three different medium on the growth and quantity of human umbilical cord mesenchymal stem cells

目的探讨三种培养基在人脐带间充质干细胞原代培养中对干细胞生长和数量的影响,从而选择更好的培养基。方法培养皿每孔脐带组织长度均为1 cm,采用同一脐带在无菌条件下收集足月分娩健康胎儿的新鲜脐带,采用组织块贴壁法在三种不同培养基中进行原代培养。分别选用:A组为低糖DMEM,B组为高糖DMEM,C组为DMEM/F12三种培养基。在7 d、10 d、15 d、20 d、25 d时间段观察三种不同培养基中细胞游出面积、细胞计数和细胞形态进行对比分析,采用SPSS 13.0重复测量方差分析进行统计学分析。结果 A组最后细胞游出面积10.42%/6孔,细胞计数(1.88±0.63)×10~4/cm;B组细胞游出面积34.32%/6孔,细胞计数(7.43±0.67)×10~4/cm;C组细胞游出面积93.13%/6孔,细胞计数(12.08±0.93)×10~4/cm。采用SPSS 13.0重复测量方差分析进行统计学分析,A组、B组、C组比较具有统计学差异(P<0.05)。结论 C组原代培养细胞游出面积明显增大,细胞计数明显增加,是一种理想的培养基。

Objective To explore the influence of three mediums on the growth and number of stem cells in the primary culture of human umbilical cord,so as to find a better medium out. Methods The length of umbilical cord tissue in each Petri dish was 1 cm. The fresh umbilical cord of healthy fetus was collected under sterile condition. The primary culture of the healthy fetus was carried out using the same umbilical cord in three different media by tissue block adherence. The selected three kinds of culture medium were as low glucose DMEM（group A）,high glucose DMEM（group B）,DMEM/F12（group C）. At 7,10,15,20,25 days after culture,the exceed area,cell count and cell morphology in three different medium were compared and analysed between three groups. Statistical analysis by SPSS 13.0 repeated measurement variance analysis is taken. Results In group A,the cell swimed out area was 10.42%/6 hole,cell count（1.88±0.63）×10~4/cm,cells in group B the swimed out area were 34.32%/6 hole,cell count（7.43±0.67）×10~4/cm;group C cells swimed out area of 93.13%/6 hole,cell count（12.08 ±0.93） ×10~4/cm,cell morphology was best. The comparison between group A and group B,group C had statistical difference（P〈0.05）. Conclution The exceed area and count of primary cultured cells in C group increase obviously,thus it could be an ideal medium.