Isolation, differentiation and characterization of vascular cells derived from human embryonic stem cells.
Isolation, differentiation and characterization of vascular cells derived from human embryonic stem cells.
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DOI：
10.1038/nprot.2010.31
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发表时间：
2010-06
期刊：
影响因子：
14.8
通讯作者：
Langer, Robert
中科院分区：
文献类型：
--
作者：
Levenberg, Shulamit;Ferreira, Lino S.;Chen-Konak, Limor;Kraehenbuehl, Thomas P.;Langer, Robert
在此，我们描述了在不同发育阶段分离人类胚胎干细胞（hESC）衍生的血管细胞的方案。这些细胞是从悬浮培养的 10 至 15 日龄人胚状体 (EB) 中分离出来的。解离后，用抗 CD34 或抗 CD31 (PECAM1) 抗体标记细胞，并通过磁激活细胞分离 (MACS) 或荧光激活细胞分选 (FACS) 从细胞混合物中分离。然后将分离的血管细胞在支持特定分化和扩张途径的培养基条件下培养。由此产生的血管细胞群含有 > 80% 的内皮样或平滑肌样细胞。假设典型的初始细胞粘附和增殖率，整个过程可以在 1.5 个月内完成。在所述条件下分离和分化的血管细胞可以构成用于修复缺血组织、制备组织工程血管移植物和设计用于药物筛选应用的细胞试剂盒的治疗应用的潜在细胞来源。
Herein, we describe a protocol for the isolation of human embryonic stem cells (hESCs)-derived vascular cells at various stages of development. The cells are isolated from 10 to 15-d-old human embryoid bodies (EBs) cultured in suspension. After dissociation, cells are labeled with anti-CD34 or anti-CD31 (PECAM1) antibody and separated from the cell mixture by magnetic-activated cell separation (MACS) or fluorescent-activated cell sorting (FACS). Isolated vascular cells are then cultured in media conditions that support specific differentiation and expansion pathways. The resulting vascular cell populations contain > 80% endothelial-like or smooth muscle-like cells. Assuming typical initial cell adhesion and proliferation rates, the entire procedure can be completed within 1.5 months. Vascular cells isolated and differentiated under the described conditions may constitute a potential cell source for therapeutic application toward repair of ischemic tissues, preparation of tissue-engineered vascular grafts and design of cellular kits for drug screening applications.
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影响因子：
56.9
作者：
Lammert, E;Cleaver, O;Melton, D
通讯作者：
Melton, D
DOI：
10.1073/pnas.1735463100
发表时间：
2003-10-28
影响因子：
11.1
作者：
Levenberg, S;Huang, NF;Langer, R
通讯作者：
Langer, R
影响因子：
20.3
作者：
Vodyanik, MA;Bork, JA;Slukvin, II
通讯作者：
Slukvin, II
DOI：
10.1073/pnas.032074999
发表时间：
2002-04-02
影响因子：
11.1
作者：
Levenberg, S;Golub, JS;Langer, R
通讯作者：
Langer, R
影响因子：
20.1
作者：
Ferreira, Lino S.;Gerecht, Sharon;Langer, Robert
通讯作者：
Langer, Robert