人脐血间充质干细胞培养方法的比较

背景：脐血中含丰富的间充质干细胞，可以作为组织工程中一种新的种子细胞来源。目的：应用两种培养基体外培养、扩增、分离纯化脐血间充质干细胞，比较其生物学特性的差异。方法：无菌条件下采取40份足月顺产的脐血，肝素抗凝。应用Ficol 密度梯度离心法分离脐血单核细胞，随机分为两组，其中20份用MesenGro人间充质干细胞培养基进行培养，20份用DMEM培养基进行培养。对比两组梭形的间充质干细胞出现时间、细胞集落出现时间、培养时间、原代细胞数量。选用生长情况良好的原代脐血间充质干细胞，用流式细胞仪检测间充质干细胞表面特异性标记表达情况。结果与结论：MesenGro组梭形的间充质干细胞平均出现时间、细胞集落平均出现时间、原代细胞平均培养时间、原代细胞平均数量为均优于DMEM组（P 〈0.01）。流式细胞仪检测显示，培养的细胞强表达间充质干细胞表面标志CD73和CD105，阳性率99.1%，不表达造血干细胞表面标志CD45和CD34，阴性率99.3%。结果提示在培养间充质干细胞的数量、形态、生长速度和培养时间诸方面，MesenGro 人间充质干细胞培养基均优于DMEM培养基。选用MesenGro人间充质干细胞培养基可以更纯、更快、更好地从脐血中培养出表达间充质干细胞表面标志的细胞。

Comparison of methods for culturing the mesenchymal stem cells from human umbilical cord blood

BACKGROUND：The umbilical cord blood is rich of mesenchymal stem cells, which can be used as a new source of seed cells in tissue engineering. OBJECTIVE：To compare two methods for culturing, expanding and purifying the mesenchymal stem cells in vitro isolated from the human umbilical cord blood. METHODS：The ful-term birth cord blood of 40 cases was col ected under sterile conditions with heparin anticoagulation. Ficol density gradient centrifugation was used to isolate the mononuclear cells from the umbilical cord blood. The cases were randomly divided into two groups according to the different culture media. Twenty cases of umbilical cord blood were cultured in MesenGro human mesenchymal stem cells culture medium （group A）, and the remaining 20 cases of umbilical cord blood were cultured in Dulbecco’s modified Eagle’s medium （group B）. The occurrence time of fusiform mesenchymal stem cells and cell colony, culture time and the number of primary cells in the two groups were compared. Cells which grew well were selected to detect the surface markers by flow cytometry. RESULTS AND CONCLUSION： The mean occurrence time of fusiform mesenchymal stem cells and cell colony, culture time and number of primary cells in group A were better than those in group B （P 〈 0.01）. The strong expression of the surface markers of mesenchymal stem cells （CD73 and CD105） was found by flow cytometry, of which the positive rate was 99.1%. No expression of the surface markers of hematopoietic stem cells （CD45 and CD34） was seen, of which the negative rate was 99.3%. The number, morphology, growth rate and culture time of umbilical cord blood mesenchymal stem cells cultured in MesenGro human mesenchymal stem cells culture medium were better than those cultured in Dulbecco’s modified Eagle’s medium. Cells cultured in MesenGro human mesenchymal stem cell culture medium can better express surface markers of mesenchymal stem cells.