Molecular stages of rapid and uniform neuralization of human embryonic stem cells

被引:30
作者
Bajpai, R.
Coppola, G.
Kaul, M.
Talantova, M.
Cimadamore, F.
Nilbratt, M.
Geschwind, D. H.
Lipton, S. A.
Terskikh, A. V.
机构
[1] Burnham Inst Med Res, Nerosci Aging & Stem Cell Res Ctr, La Jolla, CA 92037 USA
[2] Univ Calif Los Angeles, David Geffen Sch Med, Program Neurogenet, Los Angeles, CA 90095 USA
[3] Burnham Inst Med Res, Infect & Inflammatory Dis Ctr, La Jolla, CA 92037 USA
关键词
human embryonic stem cells; uniform neural differentiation; expression/coregulation analysis; NEURAL PROGENITOR CELLS; SELF-RENEWAL; IN-VIVO; DIFFERENTIATION; NEURONS; PRECURSORS; EXPRESSION; INDUCTION; ECTODERM; BRAIN;
D O I
10.1038/cdd.2009.18
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
Insights into early human development are fundamental for our understanding of human biology. Efficient differentiation of human embryonic stem cells (hESCs) into neural precursor cells is critical for future cell-based therapies. Here, using defined conditions, we characterized a new method for rapid and uniform differentiation of hESCs into committed neural precursor cells (designated C-NPCs). Dynamic gene expression analysis identified several distinct stages of ESC neuralization and revealed functional modules of coregulated genes and pathways. The first wave of gene expression changes, likely corresponding to the transition through primitive ectoderm, started at day 3, preceding the formation of columnar neuroepithelial rosettes. The second wave started at day 5, coinciding with the formation of rosettes. The majority of C-NPCs were positive for both anterior and posterior markers of developing neuroepithelium. In culture, C-NPCs became electrophysiologically functional neurons; on transplantation into neonatal mouse brains, C-NPCs integrated into the cortex and olfactory bulb, acquiring appropriate neuronal morphologies and markers. Compared to rosette-NPCs,(1) C-NPCs exhibited limited in vitro expansion capacity and did not express potent oncogenes such as PLAG1 or RSPO3. Concordantly, we never detected tumors or excessive neural proliferation after transplantation of C-NPCs into mouse brains. In conclusion, our study provides a framework for future analysis of molecular signaling during ESC neuralization. Cell Death and Differentiation (2009) 16, 807-825; doi: 10.1038/cdd.2009.18; published online 13 March 2009
引用
收藏
页码:807 / 825
页数:19
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