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A retinoic acid-induced clonal cell line derived from multipotential P19 embryonal carcinoma cells expresses smooth
R S Blank1, E A Swartz, M M Thompson
1University of Virginia School of Medicine, Department of Molecular Physiology and Biological Physics, Charlottesville, USA.
Abstract:
Despite intense interest in understanding the differentiation of vascular smooth muscle, very little is known about the cellular and molecular mechanisms that control differentiation of this cell type. Progress in this field has been hampered by the lack of an inducible in vitro system for study of the early steps of smooth muscle differentiation. In this study, we describe a model system in which multipotential mouse P19 embryonal carcinoma cells (P19s) can be induced to express multiple characteristics of differentiated smooth muscle. Treatment of P19s with retinoic acid was associated with profound changes in cell morphology and with the appearance at high frequency of smooth muscle alpha-actin-positive cells that were absent or present at extremely low frequency in parental P19s. A clonal line derived from retinoic acid-treated P19s (9E11G) stably expressed multiple characteristics of differentiated smooth muscle, including smooth muscle-specific isoforms of alpha-actin and myosin heavy chain, as well as functional responses to the contractile agonists phenylephrine, angiotensin II, ATP, bradykinin, histamine, platelet-derived growth factor (PDGF)-AA, and PDGF-BB. Additionally, 9E11G cells expressed transcripts for MHox, a muscle homeobox gene expressed in smooth, cardiac, and skeletal muscles, but not the skeletal muscle-specific regulatory factors, MyoD and myogenin. Results demonstrate that retinoic acid treatment of multipotential P19 cells is associated with formation of cell lines that stably express multiple properties of differentiated smooth muscle.(ABSTRACT TRUNCATED AT 250 WORDS)
Insights
Researchers developed a new in vitro model using mouse P19 cells to study vascular smooth muscle differentiation. Retinoic acid treatment successfully induced differentiation, creating a valuable tool for understanding these crucial cellular mechanisms.
Area of Science:
- Cell Biology
- Developmental Biology
- Muscle Physiology
Background:
- Understanding vascular smooth muscle differentiation is crucial but limited by a lack of suitable in vitro models.
- Existing models do not adequately capture the early stages of smooth muscle cell development.
Purpose of the Study:
- To establish an inducible in vitro system for studying smooth muscle cell differentiation.
- To investigate the effects of retinoic acid on P19 embryonal carcinoma cells for smooth muscle lineage induction.
Main Methods:
- Utilized multipotential mouse P19 embryonal carcinoma cells (P19s).
- Induced differentiation using retinoic acid treatment.
- Established and characterized a clonal cell line (9E11G) expressing smooth muscle markers.
Main Results:
- Retinoic acid treatment induced significant morphological changes in P19 cells.
- A high frequency of smooth muscle alpha-actin-positive cells emerged post-treatment.
- The derived 9E11G cell line stably expressed smooth muscle-specific alpha-actin and myosin heavy chain isoforms.
- 9E11G cells exhibited functional responses to various contractile agonists.
- Gene expression analysis confirmed the presence of MHox but not skeletal muscle-specific factors (MyoD, myogenin).
Conclusions:
- Retinoic acid treatment of P19 cells provides a robust model for inducing vascular smooth muscle differentiation.
- This model system facilitates the study of early cellular and molecular mechanisms governing smooth muscle development.
- The 9E11G cell line serves as a stable, differentiated smooth muscle cell source for further research.