Muscle isoactin expression during in vitro differentiation of murine embryonic stem cells

Willie A. Ng, Thomas C Doetschman, Jeffrey Robbins, James L. Lessard

Research output: Contribution to journalArticle

25 Citations (Scopus)

Abstract

Embryonic stem (ES) cells are pluripotent cells derived from mouse blastocysts. ES cells can differentiate into complex embryoid bodies (EBs) which exhibit many of the characteristics of 4-10-d embryos, including areas which rhythmically contract. The expression of the four muscle isoactins was examined in EBs by using transcript specific probes for each of the muscle actin mRNAs and selectively reactive MAbs to muscle actins. Northern blot analyses from undifferentiated ES cells and EBs after 5, 10, 15, and 20 d in suspension culture demonstrated that no muscle actin transcripts could be detected in the undifferentiated cells, whereas during differentiation, the vascular and enteric smooth muscle isoactin mRNAs were easily detected. To further define the pattern of expression, polymerase chain reaction analyses were carried out on RNA isolated from individual EBs. The data indicated that all four muscle-specific actin genes are transcribed. We also demonstrated the presence of muscle actins in at least two distinct cell populations within the EBs using selectively reactive MAbs. Fibroblast like cells exhibit significant levels of the two smooth muscle actins (vascular and enteric) localized to stress fibers. In addition, one or both of the striated muscle actins (cardiac and skeletal) are expressed in cardiomyocyte-like cells. As is the case in embryonic heart, α-smooth muscle actin and the striated muscle actin(s) are incorporated into well organized sarcomeres in these cardiomyocyte-like cells. Thus, differentiating EBs provide an in vitro system to study both striated and smooth muscle cell gene expression.

Original languageEnglish (US)
Pages (from-to)285-292
Number of pages8
JournalPediatric Research
Volume41
Issue number2
StatePublished - Feb 1997
Externally publishedYes

Fingerprint

Embryonic Stem Cells
Actins
Embryoid Bodies
Muscles
Striated Muscle
Vascular Smooth Muscle
Cardiac Myocytes
In Vitro Techniques
Stress Fibers
Messenger RNA
Sarcomeres
Blastocyst
Contracts
Northern Blotting
Smooth Muscle Myocytes
Smooth Muscle
Suspensions
Myocardium
Embryonic Structures
Fibroblasts

ASJC Scopus subject areas

  • Pediatrics, Perinatology, and Child Health

Cite this

Muscle isoactin expression during in vitro differentiation of murine embryonic stem cells. / Ng, Willie A.; Doetschman, Thomas C; Robbins, Jeffrey; Lessard, James L.

In: Pediatric Research, Vol. 41, No. 2, 02.1997, p. 285-292.

Research output: Contribution to journalArticle

Ng, Willie A. ; Doetschman, Thomas C ; Robbins, Jeffrey ; Lessard, James L. / Muscle isoactin expression during in vitro differentiation of murine embryonic stem cells. In: Pediatric Research. 1997 ; Vol. 41, No. 2. pp. 285-292.
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