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Effects of transforming growth factor-beta 1 and ascorbic acid on differentiation of human bone-marrow-derived mesenchymal stem cells into smooth muscle cell lineage

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Abstract

Bone-marrow-derived mesenchymal stem cells (MSCs) can differentiate into a variety of cell types including smooth muscle cells (SMCs). We have attempted to demonstrate that, following treatment with transforming growth factor-beta 1 (TGF-β1) and ascorbic acid (AA), human bone-marrow-derived MSCs differentiate into the SMC lineage for use in tissue engineering. Quantitative polymerase chain reaction for SMC-specific gene (α smooth muscle actin, h1-calponin, and SM22α) expression was performed on MSCs, which were cultured with various concentrations of TGF-β1 or AA. TGF-β1 had a tendency to up-regulate the expression of SMC-specific genes in a dose-dependent manner. The expression of SM22α was significantly up-regulated by 30 μM AA. We also investigated the additive effect of TGF-β1 and AA for differentiation into SMCs and compared this effect with that of other factors including platelet-derived growth factor BB (PDGF-BB). In addition to SMC-specific gene expression, SMC-specific proteins increased by two to four times when TGF-β1 and AA were used together compared with their administration alone. PDGF did not increase the expression of SMC-specific markers. MSCs cultured with TGF-β1 and AA did not differentiate into osteoblasts and adipocytes. These results suggest that a combination of TGF-β1 and AA is useful for the differentiation of MSCs into SMCs for use in tissue engineering.

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Abbreviations

AA:

ascorbic acid

AB-AM:

antibiotic-antimycotic

ALP:

alkaline phosphatase

ASMA:

alpha smooth muscle actin

BM:

basal medium

CALP:

h1-calponin

DMEM:

Dulbecco’s modified Eagle medium

ECM:

extracellular matrix

FBS:

fetal bovine serum

GAPDH:

glyceraldehyde-3-phosphate dehydrogenase

LPL:

lipoprotein lipase

MSC:

mesenchymal stem cell

PDGF:

platelet-derived growth factor

PDT:

population doubling time

PI3K:

phosphatidylinositol 3-kinase

PPARγ2:

proliferator-activated receptor-gamma2

RT-PCR:

reverse transcription and polymerase chain reaction

SMC:

smooth muscle cell

TGF-β:

transforming growth factor-beta

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Acknowledgments

We are grateful to Yosuke Murase, Shuichi Suzuki, Ryotaro Hashizume, and Taeko Komada for their technical support with the cell culture and histology and to Rina Makimura for her general management assistance.

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Authors and Affiliations

  1. Department of Clinical Cell Therapy and Tissue Engineering, Nagoya University School of Medicine, 65 Tsurumai-cho, Showa-ku, Nagoya, 466–8550, Japan

    Yuji Narita & Aika Yamawaki

  2. Division of Stem Cell Engineering, The Institute of Medical Science, The University of Tokyo, Tokyo, Japan

    Hideaki Kagami

  3. Department of Oral and Maxillofacial Surgery, Nagoya University Graduate School of Medicine, Nagoya, Japan

    Minoru Ueda

  4. Department of Cardiac Surgery, Nagoya University Graduate School of Medicine, Nagoya, Japan

    Yuji Narita & Yuichi Ueda

Authors
  1. Yuji Narita
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  2. Aika Yamawaki
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  3. Hideaki Kagami
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  4. Minoru Ueda
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  5. Yuichi Ueda
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Corresponding author

Correspondence to Yuji Narita.

Additional information

The study was supported in part by a Grant-in-Aid for Science Research (No. 17689039) from the Ministry of Education, Culture, Sports, Science and Technology of Japan.

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Narita, Y., Yamawaki, A., Kagami, H. et al. Effects of transforming growth factor-beta 1 and ascorbic acid on differentiation of human bone-marrow-derived mesenchymal stem cells into smooth muscle cell lineage. Cell Tissue Res 333, 449–459 (2008). https://doi.org/10.1007/s00441-008-0654-0

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  • DOI: https://doi.org/10.1007/s00441-008-0654-0

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