In vitro system for differentiating pluripotent neural crest cells into smooth muscle cells

Mukesh K. Jain, Matthew D. Layne, Masafumi Watanabe, Michael T. Chin, Mark W. Feinberg, Nicholas E.S. Sibinga, Chung Ming Hsieh, Shaw Fang Yet, Derek L. Stemple, Mu En Lee

Research output: Contribution to journalArticle

55 Citations (Scopus)

Abstract

The change in vascular smooth muscle cells (SMC) from a differentiated to a dedifferentiated state is the critical phenotypic response that promotes occlusive arteriosclerotic disease. Despite its importance, research into molecular mechanisms regulating smooth muscle differentiation has been hindered by the lack of an in vitro cell differentiation system. We identified culture conditions that promote efficient differentiation of Monc- 1 pluripotent neural crest cells into SMC. Exclusive Monc-1 to SMC differentiation was indicated by cellular morphology and time-dependent induction of the SMC markers smooth muscle α-actin, smooth muscle myosin heavy chain, calponin SM22α, and APEG-1. The activity of the SM22α promoter was low in Monc-1 cells. Differentiation of these cells into SMC caused a 20- 30-fold increase in the activity of the wild-type SM22α promoter and that of a hybrid promoter containing three copies of the CArG element. By gel mobility shift analysis, we identified new DNA-protein complexes in nuclear extracts prepared from differentiated Monc-1 cells. One of the new complexes contained serum response factor. This Monc-1 to SMC model should facilitate the identification of nodal regulators of smooth muscle development and differentiation.

Original languageEnglish (US)
Pages (from-to)5993-5996
Number of pages4
JournalJournal of Biological Chemistry
Volume273
Issue number11
DOIs
StatePublished - Mar 13 1998
Externally publishedYes

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Neural Crest
Smooth Muscle Myocytes
Muscle
Cells
Smooth Muscle
Cell Differentiation
Serum Response Factor
Smooth Muscle Myosins
Myosin Heavy Chains
Muscle Development
Electrophoretic Mobility Shift Assay
Vascular Smooth Muscle
In Vitro Techniques
Actins
DNA
Identification (control systems)
Gels
Research
Proteins

ASJC Scopus subject areas

  • Biochemistry

Cite this

In vitro system for differentiating pluripotent neural crest cells into smooth muscle cells. / Jain, Mukesh K.; Layne, Matthew D.; Watanabe, Masafumi; Chin, Michael T.; Feinberg, Mark W.; Sibinga, Nicholas E.S.; Hsieh, Chung Ming; Yet, Shaw Fang; Stemple, Derek L.; Lee, Mu En.

In: Journal of Biological Chemistry, Vol. 273, No. 11, 13.03.1998, p. 5993-5996.

Research output: Contribution to journalArticle

Jain, MK, Layne, MD, Watanabe, M, Chin, MT, Feinberg, MW, Sibinga, NES, Hsieh, CM, Yet, SF, Stemple, DL & Lee, ME 1998, 'In vitro system for differentiating pluripotent neural crest cells into smooth muscle cells', Journal of Biological Chemistry, vol. 273, no. 11, pp. 5993-5996. https://doi.org/10.1074/jbc.273.11.5993
Jain, Mukesh K. ; Layne, Matthew D. ; Watanabe, Masafumi ; Chin, Michael T. ; Feinberg, Mark W. ; Sibinga, Nicholas E.S. ; Hsieh, Chung Ming ; Yet, Shaw Fang ; Stemple, Derek L. ; Lee, Mu En. / In vitro system for differentiating pluripotent neural crest cells into smooth muscle cells. In: Journal of Biological Chemistry. 1998 ; Vol. 273, No. 11. pp. 5993-5996.
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