| 贾 静,余 俊,王少帅,张婧怡,冯 玲.人脐带血中内皮克隆形成细胞的分离和培养[J].现代生物医学进展英文版,2018,(10):1812-1815. |
| 人脐带血中内皮克隆形成细胞的分离和培养 |
| Isolation and Primary Culture of Endothelial Colony Forming Cells Derived from Human Umbilical Cord Blood |
| Received:January 03, 2018 Revised:January 27, 2018 |
| DOI：10.13241/j.cnki.pmb.2018.10.003 |
| 中文关键词: 内皮克隆形成细胞 脐带血 细胞分离和培养 |
| 英文关键词: Endothelial colony-forming cells (ECFCs) Cord blood Cell isolation and culture |
| 基金项目:国家重点研究发展计划项目(2016YFC1000405) |
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| 中文摘要: |
| 摘要 目的：脐带血来源的内皮克隆形成细胞具有高度增殖、自我更新和血管生成的能力，是再生医学和母胎医学领域研究的热点。本研究旨在提出一种有效的、可靠的人脐带血中内皮克隆形成细胞分离和培养方法。方法：取抗凝脐血，采用密度梯度离心法分离单核细胞并进行贴壁培养，在细胞克隆形成但未融合成片时，进行单克隆的挑取再培养，最后对所得细胞进行表型和功能的鉴定。结果：所获细胞增殖能力强，形态单一，呈鹅卵石样，连续传代8次，未见细胞性状改变。接种到基质胶上，细胞可连接为管状、网状结构。能摄取培养液中的低密度脂蛋白并结合荆豆凝集素，细胞免疫荧光显示其表达典型的内皮细胞标志性分子CD31、eNOS、VE-Cadherin、vWF。结论：通过本研究的方法，我们从脐带血中获得足够数量和高纯度的内皮克隆形成细胞，这为进一步的实验研究提供了可靠的基础。 |
| 英文摘要: |
| ABSTRACT Objective: Umbilical cord blood-derived endothelial colony forming cells(ECFCs) have high proliferation potential, self-renewal and vessel forming capacity. It is well studied in the field of regenerative and maternal-fetal medicine. This study aims to propose an effective and reliable method for ECFCs isolation and culture from umbilical cord blood. Methods: Heparin is used as antico- agulant, density gradient centrifugation and adherent culture is used for ECFCs isolation. When clones are formed but not fused, mono- clonal is selected and passaged. Finally, the obtained cells are subjected to phenotypic and functional characterization. Results: The ob- tained cells exhibit high proliferation ability , presented as cobblestone-like appearance and can form tubular structure. They can intake ac-LDL, combine Ulex-lectin and express the typical molecular markers of endothelial cells(namely CD31, eNOS, VE-Cadherin, vWF). Conclusion: By this efficient isolation method, sufficient and high-quality of ECFCs from cord blood can be obtained, providing a reli- able basis for further experimental research. |
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