Dataset Information

Laminin-221-derived recombinant fragment facilitates isolation of cultured skeletal myoblasts.

ABSTRACT:

Introduction

Laminin is a major component of the basement membrane, containing multiple domains that bind integrin, collagen, nidogen, dystroglycan, and heparan sulfate. Laminin-221, expressed in skeletal and cardiac muscles, has strong affinity for the cell-surface receptor, integrin α7X2β1. The E8 domain of laminin-221, which is essential for cell integrin binding, is commercially available as a purified recombinant protein fragment. In this study, recombinant E8 fragment was used to purify primary rodent myoblasts. We established a facile and inexpensive method for primary myoblast culture exploiting the high affinity binding of integrin α7X2β1 to laminin-221.

Methods

Total cell populations from dissociated muscle tissue were enzymatically digested and seeded onto laminin-221 E8 fragment-coated dishes. The culture medium containing non-adherent floating cells was removed after 2-hour culture at 37 °C. The adherent cells were subjected to immunofluorescence staining of desmin, differentiation experiments, and gene expression analysis.

Results

The cells obtained were 70.3 ± 5.49% (n = 5) desmin positive in mouse and 67.7 ± 1.65% (n = 3) in rat. Immunofluorescent staining and gene expression analyses of cultured cells showed phenotypic traits of myoblasts.

Conclusion

This study reports a novel facile method for primary culture of myoblasts obtained from mouse and rat skeletal muscle by exploiting the high affinity of integrin α7X2β1 to laminin-221.

SUBMITTER: Kihara Y

PROVIDER: S-EPMC9111930 | biostudies-literature | 2022 Jun

REPOSITORIES: biostudies-literature

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Publications

Laminin-221-derived recombinant fragment facilitates isolation of cultured skeletal myoblasts.

Kihara Yuki Y Homma Jun J Takagi Ryo R Ishigaki Keiko K Nagata Satoru S Yamato Masayuki M

Regenerative therapy 20220512

<h4>Introduction</h4>Laminin is a major component of the basement membrane, containing multiple domains that bind integrin, collagen, nidogen, dystroglycan, and heparan sulfate. Laminin-221, expressed in skeletal and cardiac muscles, has strong affinity for the cell-surface receptor, integrin α7X2β1. The E8 domain of laminin-221, which is essential for cell integrin binding, is commercially available as a purified recombinant protein fragment. In this study, recombinant E8 fragment was used to p ...[more]

PMID: 35620637

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Dataset Information

Laminin-221-derived recombinant fragment facilitates isolation of cultured skeletal myoblasts.

Introduction

Methods

Results

Conclusion

Publications

Laminin-221-derived recombinant fragment facilitates isolation of cultured skeletal myoblasts.

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