| 摘要: |
| ［摘要］ 目的 脐带血间充质干细胞为组织工程种子细胞领域研究热点，但目前仍未得到安全、高效的脐带血间充质干细胞分离培养体系，该实验研究旨在探讨脐带血间充质干细胞的适宜培养条件。方法 无菌条件下取足月正常新生儿脐带血，随机分为高糖DMEM组、低糖DMEM组以及脐带血间充质干细胞组。分离提取的脐带血单个核细胞分别以1×104、1×105和1×106个细胞/ml的接种密度接种于脐带血间充质干细胞培养基中，观察3种培养基及不同接种密度培养下的细胞贴壁生长情况，并用流式细胞技术分析细胞的表面抗原。结果 高糖DMEM组未见到贴壁生长的间充质干细胞，低糖DMEM组及脐带血间充质干细胞组均可见到贴壁生长的梭形成纤维样细胞（P<0.05），在不同的接种密度条件下，1×105组及1×106组接种密度可见到成纤维样贴壁细胞，1×105组得到的细胞数量及细胞生长情况更好，1×104组未见到细胞贴壁生长。结论 分离提取得到的脐带血单个核细胞在T25培养瓶中最佳接种密度为1×105/ml，最适合培养基为脐带血间充质干细胞培养基。 |
| 关键词: 脐带血 间充质干细胞 分离培养 |
| DOI：10.3969/j.issn.1674-3806.2015.09.02 |
| 分类号:Q 813 |
| 基金项目:广西医疗卫生重点科研课题（编号：重2011013) |
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| Different culture media, and inoculation density cultivate ectomesenchymal stem cells, umbilical cord blood |
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XUE Xing-ying, KE Ru-xiang, QIN Jin-ding, et al.
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The Fourth Department of Orthopaedics, the Second Hospital of Fuzhou City Affiliated to Xiamen University, Fuzhou 350000, China
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| Abstract: |
| ［Abstract］ Objective To explore the appropriate culture conditions of umbilical cord blood mesenchymal stem cells.Methods The sterile cord blood from normal full-term newborns was randomly divided into three groups: the high sugar DMEM group, the low sugar DMEM group and the umbilical cord blood mesenchymal stem cells between groups. Extractions of umbilical cord blood mononuclear cells were inoculated in umbilical cord blood mesenchymal stem cells in the culture media by the density of 1×105, 1×104, and 1×106 cells/ml respectively. The three kinds of media and different inoculation density culture adherent cell growth situations were observed and analysis of cell surface antigen was performed using flow cytometry technique.Results Ectomesenchymal stem cells of sugar DMEM group did not grow on the sidewall, while the growth of the spindle formation fiber sample cells was found on the sidewall in low sugar DMEM group and umbilical cord blood mesenchymal stem cell(P<0.05). Of different inoculation densities, cells grew best at the density of 1×105, followed by 1×106. Cells failed to grow at the density of 1×104.Conclusion Umbilical cord blood mononuclear cells in T25 culture bottle grow best at the inoculation density of 1×105/ml, which is the most suitable culture medium for umbilical cord blood mesenchymal stem cell. |
| Key words: Umbilical cord blood Mesenchymal stem cell Isolated culture |
