体外模拟干细胞微环境培养扩增脐血及脐带基质源性间充质干细胞**

背景：建立稳健的扩增体系,在体外扩增时最大限度地获得治疗剂量的干细胞数,同时保存干细胞的特性是临床实验室亟待解决的问题。目的：建立体外模拟干细胞微环境细胞外基质扩增脐血间充质干细胞及脐带间充质干细胞的方法,并与传统的2-D培养体系比较。方法：建立体外模拟干细胞微环境细胞外基质,体外分离脐血间充质干细胞及脐带间充质干细胞后分别接种到细胞外基质及传统的2-D塑料培养体系,分别从细胞计数及细胞表面标志物的变化,评估两种扩增体系对脐血间充质干细胞及脐带间充质干细胞体外扩增的优劣。结果与结论：使用骨髓源性的细胞外基质培养体系单位时间脐血间充质干细胞及脐带间充质干细胞产量是2-D体系的4~6倍,并且流式细胞仪检测显示细胞外基质体系能更好地保持干细胞的表面标记。因此,3-D较2-D培养系更接近生理环境,已建立的骨髓源性细胞外基质培养体系能保持间充质干细胞的特性,为短时间内快速获取更大数目同质、高活性的间充质干细胞提供了可能。关键词：脐血间充质干细胞;脐带间充质干细胞;细胞外基质;2-D培养板;培养扩增doi:10.3969/j.issn.1673-8225.2012.10.011

Simulation of stem cell microenvironment in vitro to culture and expand cord and umbilical cord mesenchymal stem cells

BACKGROUND: The future of cell therapy requires high purity of mesenchymal stem cells (MSCs) in order to clearly determine the therapeutic dose and dose-response relationship. Establishing a robust amplification system in vitro to obtain the therapeutic dose of stem cells while preserving the characteristics of stem cells is the urgent of clinical laboratories. OBJECTIVE: To simulate a microenvironment for stem cells in vitro to harvest and expand umbilical cord blood-derived and cord-derived MSCs and to compare with conventional two-dimensional culture system.METHODS: The umbilical cord blood-derived and cord-derived MSCs were inoculated in the conventional two-dimensional plastic culture system and extracellular matrix, respectively. The two amplification systems for in vitro large-scale expansion of umbilical cord blood-derived and cord-derived MSCs were evaluated from the points of cell counts and surface markers.RESULTS AND CONCLUSION: The production of umbilical cord blood-derived and cord-derived MSCs in the bone marrow-derived extracellular matrix culture system was 4-6 times of that in the conventional two-dimensional culture system. FACS analysis showed that extracellular matrix system better maintained the surface marker of stem cells. Therefore, such a three-dimensional culture system is much closer to physical environment than the two-dimensional culture system. The established bone marrow-derived extracellular matrix culture system can maintain the characteristics of mesenchymal stem cells, which provides access to more homogeneous MSCs of high activity within a short time period.