Citation

Derivation of Neural Crest from Mouse Embryonic Stem Cells.

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Abstract:

Unlike the skeletal elements in the appendicular or axial skeleton, much of the facial skeleton is formed by cranial neural crest cells (NCC). Although NCC have been isolated from adult tissues, there are major challenges in utilization of adult stem cells in regenerative medicine. The high proliferative activity and pluripotency of embryonic stem (ES) cells provide a promising tool for regenerative medicine and clinical applications in transplantation therapy. Objective: To examine the formation of NCC-like cells using mouse ES cell line, 46C in which the expression of green fluorescent protein (GFP) is under the control of Sox1 promoter, the earliest known specific marker for neuroectoderm. Methods: Undifferentiated 46C mouse ES cells were cultured without a feeder layer on gelatin-coated dishes in serum-free neuronal differentiation media containing N2 and B27 for various time points. The formation of various cell types including NCC-like cells was examined by immunocytochemistry, RT-PCR and fluorescence-activated cell sorting (FACS). Results: Undifferentiated 46C cells expressed Oct4 but not Sox1-GFP. Sox1-GFP+ cells appeared as rosettes at day 3 and increased over time coincident with down- regulation of Oct4. The NCC-like cells characterized by the expression of p75, c-Kit and snail formed in close proximity to Sox1-GFP+ cells. The percentage of NCC-like cells increased with time in culture. The formation of neurons at day 6 adjacent to Sox1-GFP+/P75+ cells was evident by immunostaining with Tuj1+. Primitive endoderm identified by the expression of Troma-1 appeared at day 6 in these cultures. Cells stained with Troma-1 were excluded from Sox1-GFP+ and P75+ cells. Conclusion: The Sox1–GFP monolayer culture provide a powerful system for examining the underlying mechanism for effective generation of NCC from ES cells. Supported by Grant 06SCC04 from CT Stem Cell Research Fund.
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Association:
Name: Connecticut's Stem Cell Research International Symposium
URL:
http://stemconn.org


Citation:
URL: http://citation.allacademic.com/meta/p319342_index.html
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MLA Citation:

Rodgers, Barbara., Fu, Yu., Roberts, Chanda., Balic, Anamaria., Wang, Y-H., Aguila, Hector. and Mina, Mina. "Derivation of Neural Crest from Mouse Embryonic Stem Cells." Paper presented at the annual meeting of the Connecticut's Stem Cell Research International Symposium, Omni Hotel, New Haven, CT, Mar 23, 2009 <Not Available>. 2014-11-29 <http://citation.allacademic.com/meta/p319342_index.html>

APA Citation:

Rodgers, B. , Fu, Y. , Roberts, C. , Balic, A. , Wang, Y. , Aguila, H. L. and Mina, M. , 2009-03-23 "Derivation of Neural Crest from Mouse Embryonic Stem Cells." Paper presented at the annual meeting of the Connecticut's Stem Cell Research International Symposium, Omni Hotel, New Haven, CT <Not Available>. 2014-11-29 from http://citation.allacademic.com/meta/p319342_index.html

Publication Type: Poster
Review Method: Peer Reviewed
Abstract: Unlike the skeletal elements in the appendicular or axial skeleton, much of the facial skeleton is formed by cranial neural crest cells (NCC). Although NCC have been isolated from adult tissues, there are major challenges in utilization of adult stem cells in regenerative medicine. The high proliferative activity and pluripotency of embryonic stem (ES) cells provide a promising tool for regenerative medicine and clinical applications in transplantation therapy. Objective: To examine the formation of NCC-like cells using mouse ES cell line, 46C in which the expression of green fluorescent protein (GFP) is under the control of Sox1 promoter, the earliest known specific marker for neuroectoderm. Methods: Undifferentiated 46C mouse ES cells were cultured without a feeder layer on gelatin-coated dishes in serum-free neuronal differentiation media containing N2 and B27 for various time points. The formation of various cell types including NCC-like cells was examined by immunocytochemistry, RT-PCR and fluorescence-activated cell sorting (FACS). Results: Undifferentiated 46C cells expressed Oct4 but not Sox1-GFP. Sox1-GFP+ cells appeared as rosettes at day 3 and increased over time coincident with down- regulation of Oct4. The NCC-like cells characterized by the expression of p75, c-Kit and snail formed in close proximity to Sox1-GFP+ cells. The percentage of NCC-like cells increased with time in culture. The formation of neurons at day 6 adjacent to Sox1-GFP+/P75+ cells was evident by immunostaining with Tuj1+. Primitive endoderm identified by the expression of Troma-1 appeared at day 6 in these cultures. Cells stained with Troma-1 were excluded from Sox1-GFP+ and P75+ cells. Conclusion: The Sox1–GFP monolayer culture provide a powerful system for examining the underlying mechanism for effective generation of NCC from ES cells. Supported by Grant 06SCC04 from CT Stem Cell Research Fund.


Similar Titles:
Generation and Transplantation of Human Embryonic Stem Cell Derived Neural Precursor Cells

An Efficient Protocol for Generating Neural Stem Cells (NSCs) from Human Embryonic Stem Cells

Inhibiting Tumor Formation and Promoting Integration of Embryonic Stem Cell Derived Neural Stem Cell Transplants into the Mouse Hippocampus


 
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