摘要
背景:胎盘间充质干细胞能够为细胞治疗提供理想的种子细胞,但其培养未曾使用过胰酶冷消化法,故对此方法进行探索,并与组织块法进行比较,从中筛选出一种方便易行、培养成功率更高的方法。目的:观察胰酶冷消化法与组织块法分离培养的人胎盘间充质干细胞的生物学特性。方法:采用胰酶冷消化法和组织块法从人胎盘中分离、纯化和传代培养人胎盘间充质干细胞(n=6)。记录胎盘间充质干细胞首次出现时间及原代培养周期,绘制第3代细胞生长曲线,流式细胞仪分析第3代细胞表面标志,检测其向成神经及成脂方向诱导分化能力。结果与结论:(1)使用上述2种培养方法均可获得胎盘间充质干细胞,胰酶冷消化法首次出现贴壁细胞时间早于组织块法(P <0.05),原代培养周期短于组织块法(P <0.05);(2)生长曲线提示在进入平台期后的各个时间点胰酶冷消化法培养的细胞数量明显多于组织块法(P<0.05);(3)2种细胞具有均一的细胞表型,均表达CD29,CD105,不表达CD34,CD45;(4)2种方法培养的胎盘间充质干细胞经诱导后均具有成神经及成脂分化潜能;(5)上述结果表明,对于胎盘间充质干细胞的培养而言,胰酶冷消化法具有明显优势。
BACKGROUND:Placental mesenchymal stem cells can provide ideal seed cells for cell therapy,but trypsin cold digestion has not been used for cell culture.Therefore,we explored this method and compared it with the tissue explant method to screen a more convenient and more successful culture method.OBJECTIVE:To observe the biological features of human placental mesenchymal stem cells isolated by trypsin cold digestion and tissue explant method.METHODS:We isolated,purified,and subcultured mesenchymal stem cells(n=6)from human placenta by using trypsin cold digestion method and tissue explant method.The first appearance time and primary culture period of placental mesenchymal stem cells isolated using two methods were recorded.The growth curves of passage 3 cells from two culture methods were generated.Flow cytometry was used to analyze the expression of surface markers of passage 3 cells isolated using the two culture methods,and to study the neural and adipogenic differentiation of the cells.RESULTS AND CONCLUSION:Placental mesenchymal stem cells could be obtained using both culture methods.Adherent cells in the trypsin cold digestion group appeared significantly earlier than those in the tissue explant group(P<0.05),and the primary culture period was also significantly shorter in the trypsin cold digestion method(P<0.05).The growth curve of passage 3 cells showed a higher cell number at plateau growth using trypsin cold digestion than tissue explant method(P<0.05).The cells isolated using two culture methods were positive for CD29 and CD105,but negative for CD34 and CD45,indicating the same cell phenotypes.After the induction,placental mesenchymal stem cells cultured by both methods had the potential to differentiate into nerve cells and adipocytes.Therefore,both culture methods can produce placental mesenchymal stem cells,but trypsin cold digestion method is more suitable for the culture of this kind of cells as compared with the tissue explant method.
作者
张飞
王武
李慧娟
刘亚飞
张宝刚
裴富强
邵青伟
武忠炎
Zhang Fei;Wang Wu;Li Hui-juan;Liu Ya-fei;Zhang Bao-gang;Pei Fu-qiang;Shao Qing-wei;Wu Zhong-yan(Xi’an Aerospace General Hospital,Xi’an 710100,Shaanxi Province,China;Department of Orthopedics,Fifth Affiliated Hospital of Xinjiang Medical University,Urumqi 830001,Xinjiang Uygur Autonomous Region,China;Department of Bone Repair and Remodeling,the Fifth Affiliated Hospital of Xinjiang Medical University,Urumqi 830001,Xinjiang Uygur Autonomous Region,China;Xi’an XD Group Hospital,Xi’an 710100,Shaanxi Province,China)
出处
《中国组织工程研究》
CAS
北大核心
2018年第33期5292-5296,共5页
Chinese Journal of Tissue Engineering Research
基金
新疆维吾尔自治区自然科学基金项目(2016D01C239)。
