Directed Differentiation of Human Embryonic Stem Cells to Neural Crest Stem Cells, Functional Peripheral Neurons, and Corneal Keratocytes

被引：21

作者：

Zhu, Qian ^{[1
]}

Li, Mingming ^{[1
]}

Yan, Chuan ^{[2
,3
]}

Lu, Qiqi ^{[1
]}

Wei, Shunhui ^{[4
]}

Gao, Rong ^{[1
]}

Yu, Mengfei ^{[5
]}

Zou, Yu ^{[1
]}

Sriram, Gopu ^{[1
,6
]}

Tong, Huei J. ^{[1
]}

Hunziker, Walter ^{[4
]}

Seneviratne, Chaminda J. ^{[1
]}

Gong, Zhiyuan ^{[2
,3
]}

Olsen, Bjorn R. ^{[7
,8
]}

Cao, Tong ^{[1
,3
,9
]}

机构：

[1] Natl Univ Singapore, Fac Dent, Singapore, Singapore

[2] Natl Univ Singapore, Dept Biol Sci, Singapore, Singapore

[3] Natl Univ Singapore, Natl Univ Singapore Grad Sch Integrat Sci & Engn, Singapore, Singapore

[4] Inst Mol & Cell Biol, Epithelial Cell Biol Lab, Singapore, Singapore

[5] Zhejiang Univ, Affiliated Stomatol Hosp, Hangzhou 310003, Zhejiang, Peoples R China

[6] Agcy Sci Technol & Res, Inst Med Biol, Singapore, Singapore

[7] Harvard Med Sch, Boston, MA 02115 USA

[8] Harvard Sch Dent Med, Boston, MA 02115 USA

[9] Natl Univ Singapore, NUSTEP, Inst Life Sci, Singapore, Singapore

来源：

BIOTECHNOLOGY JOURNAL | 2017年 / 12卷 / 12期

关键词：

differentiation; human embryonic stem cells; keratocyte; neural crest stem cells; peripheral neuron; SENSORY NEURONS; GROWTH-FACTOR; INDUCTION; PROLIFERATION; REGENERATION; MAINTENANCE; INHIBITION; ACTIVATION; GENERATION; DERIVATION;

D O I：

10.1002/biot.201700067

中图分类号：

Q5 [生物化学];

学科分类号：

071010 ; 081704 ;

摘要：

Neural crest stem cells (NCSCs) are a transient and multipotent cell population giving rise to various cell types with clinical importance. Isolation of human NCSCs is extremely challenging that limits our knowledge about neural crest development and application. Here, a defined protocol to efficiently direct human embryonic stem cells (hESCs) to NCSCs and multiple neural crest lineages is presented. A unique combination of small molecule inhibitors and growth factors is employed to generate NCSCs from hESCs through a neuroectoderm stage. The self-renewal and multipotent capacities of hESC-derived NCSCs are assessed subsequently. In the feeder-free system, hESC-derived NCSCs (P75(+)/HNK1(+)/AP2(+)/PAX6(-)) in high purity are efficiently generated following neuroectodermal restriction. They can be propagated and differentiated toward multiple neural crest lineages in vitro, such as functional peripheral neurons (-tubulin III+/peripherin(+)), mesenchymal stem cells (CD73(+)CD90(+)CD105(+)), and corneal keratocytes (keratocan(+)). The in vivo developmental potential of hESC-derived NCSCs is confirmed using zebrafish embryos. This report is the first demonstration of efficient differentiation of hESCs into corneal keratocytes as a monolayer in a feeder-free system. Considering the high efficacy of NCSC generation, this new method will be a useful tool for future clinical organ repair and regeneration, such as peripheral nerve regeneration and corneal repair.

引用

页数：11

共 49 条

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