NDLI: Genetic manipulation of neural progenitors derived from human embryonic stem cells.

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Genetic manipulation of neural progenitors derived from human embryonic stem cells.

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Genetic manipulation of neural progenitors derived from human embryonic stem cells.

Content Provider	World Health Organization (WHO)-Global Index Medicus
Author	Dhara, Sujoy K. Gerwe, Brian A. Majumder, Anirban Dodla, Mahesh C. Boyd, Nolan L. Machacek, David W. Hasneen, Kowser Stice, Steven L.
Description	Country affiliation: United States Author Affiliation: Dhara SK ( Regenerative Bioscience Center, University of Georgia, Athens, Georgia 30602, USA.)
Abstract	Human embryonic stem cell-derived neural progenitors (NP) present an important tool for understanding human development and disease. Optimal utilization of NP cells, however, requires an enhanced ability to monitor these cells in vitro and in vivo. Here we report production of the first genetically modified self-renewing human embryonic stem cell-derived NP cells that express fluorescent proteins under constitutive as well as lineage-specific promoters, enabling tracking and monitoring of cell fate. Nucleofection, transfection, and lentiviral transduction were compared for optimal gene delivery to NP cells. Transduction was most efficient in terms of transgene expression (37%), cell viability (39%), and long-term reporter expression (>3 months). Further, the constitutive gene promoters, cytomegalovirus, elongation factor 1alpha, and ubiquitin-C, exhibited comparable silencing (20-30%) in NP cells over a 2-month period, suggesting their suitability for long-term reporter expression studies. Transduced NP cells maintained their progenitor state and differentiation potential, as demonstrated by expression of endogenous NP markers and neuronal markers after differentiation. We also detected reporter expression in astrocytes generated from NP cells transduced with an astrocyte-specific gene promoter, glial fibrillary acidic protein, demonstrating the usefulness of this approach. The genetically manipulated NP cells described here offer great potential for live cell-tracking experiments, and a similar approach can as well be used for expression of proteins other than reporters.
File Format	HTM / HTML
ISSN	21524947
e-ISSN	21524955
DOI	10.1089/ten.tea.2009.0155
Journal	Tissue Engineering Part A
Issue Number	11
Volume Number	15
Language	English
Publisher	Mary Ann Liebert, Inc.
Publisher Date	2009-11-01
Publisher Place	United States
Access Restriction	Open
Subject Keyword	Discipline Biotechnology Embryonic Stem Cells Cytology Physiology Genetic Enhancement Neurons Cell Differentiation Genetics Cell Proliferation Cell Survival Cells, Cultured Research Support, N.i.h., Extramural Research Support, Non-u.s. Gov't Research Support, U.s. Gov't, Non-p.h.s.
Content Type	Text
Resource Type	Article

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1	Ministry of Education (GoI), Department of Higher Education	Sanctioning Authority	https://www.education.gov.in/ict-initiatives
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3	National Digital Library of India Office, Indian Institute of Technology Kharagpur	The administrative and infrastructural headquarters of the project	Dr. B. Sutradhar bsutra@ndl.gov.in
4	Project PI / Joint PI	Principal Investigator and Joint Principal Investigators of the project	Dr. B. Sutradhar bsutra@ndl.gov.in Prof. Saswat Chakrabarti will be added soon
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