In vitro generation of three-dimensional substrate-adherent embryonic stem cell-derived neural aggregates for application in animal models of neurological disorders.
In vitro generation of three-dimensional substrate-adherent embryonic stem cell-derived neural aggregates for application in animal models of neurological disorders.
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体外生成三维基底粘附胚胎干细胞衍生的神经聚集体，用于神经系统疾病动物模型
DOI：
10.1002/9780470151808.sc02d11s21
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发表时间：
2012
影响因子：
--
通讯作者：
Schachner
中科院分区：
文献类型：
--
作者：
Hargus;Dihné;Bernreuther;Schachner
体外分化的胚胎干 (ES) 细胞是细胞替代疗法的有用来源，但转化方法的效率和安全性高度依赖于将 ES 细胞在体外定向分化为所需细胞类型的优化方案。此外，三维 ES 细胞衍生结构的移植而不是单细胞悬浮液可以通过为植入细胞提供有益的微环境来改善移植物的存活和功能。为此，我们开发了一种新方法，可以有效地将小鼠 ES 细胞分化为神经聚集体，该神经聚集体主要由（> 90%）有丝分裂后神经元、神经祖细胞和放射状胶质细胞样细胞组成。当移植到成年小鼠兴奋性毒性损伤的纹状体中时，这些基底粘附的胚胎干细胞衍生的神经聚集体（SENA）比移植的ES细胞衍生的神经细胞的单细胞悬浮液显示出显着的优势，包括提高GABA能神经元的存活率，增加细胞迁移，并显着降低畸胎瘤形成的风险。此外，SENA 移植到帕金森病和脊髓损伤的动物模型后可介导功能改善。本单元详细描述了如何在体外有效地从小鼠 ES 细胞中提取 SENA 以及如何分离 SENA 进行移植。此外，还提出了将 SENA 成功植入亨廷顿病、帕金森病和脊髓损伤动物模型的方法，以研究干细胞衍生的神经聚集体在体内疾病背景下的作用。协议。干细胞生物学。 21：2D.11.1-2D.11.19。 © 2012 John Wiley & Sons, Inc. 版权所有
In vitro−differentiated embryonic stem (ES) cells comprise a useful source for cell replacement therapy, but the efficiency and safety of a translational approach are highly dependent on optimized protocols for directed differentiation of ES cells into the desired cell types in vitro. Furthermore, the transplantation of three‐dimensional ES cell−derived structures instead of a single‐cell suspension may improve graft survival and function by providing a beneficial microenvironment for implanted cells. To this end, we have developed a new method to efficiently differentiate mouse ES cells into neural aggregates that consist predominantly (>90%) of postmitotic neurons, neural progenitor cells, and radial glia−like cells. When transplanted into the excitotoxically lesioned striatum of adult mice, these substrate‐adherent embryonic stem cell−derived neural aggregates (SENAs) showed significant advantages over transplanted single‐cell suspensions of ES cell−derived neural cells, including improved survival of GABAergic neurons, increased cell migration, and significantly decreased risk of teratoma formation. Furthermore, SENAs mediated functional improvement after transplantation into animal models of Parkinson's disease and spinal cord injury. This unit describes in detail how SENAs are efficiently derived from mouse ES cells in vitro and how SENAs are isolated for transplantation. Furthermore, methods are presented for successful implantation of SENAs into animal models of Huntington's disease, Parkinson's disease, and spinal cord injury to study the effects of stem cell−derived neural aggregates in a disease context in vivo.Curr. Protoc. Stem Cell Biol. 21:2D.11.1‐2D.11.19. © 2012 by John Wiley & Sons, Inc.
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影响因子：
3.7
作者：
Cui YF;Xu JC;Hargus G;Jakovcevski I;Schachner M;Bernreuther C
通讯作者：
Bernreuther C
DOI：
10.1073/pnas.97.11.6126
发表时间：
2000-05-23
影响因子：
11.1
作者：
Liu, S;Qu, Y;McDonald, JW
通讯作者：
McDonald, JW
影响因子：
14.5
作者：
Buehnemann, Claudia;Scholz, Andreas;Dihne, Marcel
通讯作者：
Dihne, Marcel
DOI：
10.1073/pnas.78.12.7634
发表时间：
1981-01-01
期刊：
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA-BIOLOGICAL SCIENCES
影响因子：
--
作者：
MARTIN, GR
通讯作者：
MARTIN, GR
DOI：
10.1073/pnas.0511153103
发表时间：
2006-02-21
影响因子：
11.1
作者：
Martinat, C;Bacci, JJ;Abeliovich, A
通讯作者：
Abeliovich, A