Optimization of human skeletal muscle precursor cell culture and myofiber formation in vitro

Daniel Eberli, Shay Soker, Anthony Atala, James J. Yoo

Research output: Contribution to journalArticle

41 Citations (Scopus)

Abstract

Muscle bioengineering is proposed as a treatment option for various conditions requiring restoration of muscle function. In order to allow for rapid clinical translation culture conditions have to be optimized for human application. The optimal isolation and culture technique should be able to support cell growth and differentiation using defined media only. Therefore, we have evaluated alternative culture conditions to determine the optimal growth condition for the engineering of human skeletal muscle. In this research, we present protocols for consistent isolation and growth of human muscle precursor cells (MPCs). MPCs were grown from human biopsies and expanded in culture using defined media and collagen coated dishes only. The best results were achieved using a one-step pre-plating and by supplementing the growth medium with insulin, dexamethasone, human basic fibroblast growth factor (hFGF) and human epithelial growth factor (hEGF). Detailed cell characterization using fluorescence-activated cell-sorting analysis and morphological analysis at different passages were performed. Further, the applicability of these cells for tissue engineering purposes was assessed by measuring expansion potential, formation of myofibers and fused myotubes. We have established a culture technique for human MPCs that allows for reliable cell growth and expansion using collagen coated dishes and defined media only. Cell characterization demonstrated a muscle phenotype and the ability to form myofibers in vitro.

Original languageEnglish
Pages (from-to)98-103
Number of pages6
JournalMethods
Volume47
Issue number2
DOIs
StatePublished - Feb 1 2009
Externally publishedYes

Fingerprint

Cell culture
Muscle Cells
Muscle
Skeletal Muscle
Cell Culture Techniques
Myoblasts
Growth
Culture Techniques
Muscles
Cell growth
Collagen
Cell Engineering
Bioengineering
Aptitude
Human Engineering
Skeletal Muscle Fibers
Fibroblast Growth Factor 2
Tissue Engineering
Biopsy
Bioelectric potentials

Keywords

  • Clinical translation
  • Human tissue engineering
  • Muscle precursor cells
  • Muscle tissue

ASJC Scopus subject areas

  • Molecular Biology
  • Biochemistry, Genetics and Molecular Biology(all)

Cite this

Optimization of human skeletal muscle precursor cell culture and myofiber formation in vitro. / Eberli, Daniel; Soker, Shay; Atala, Anthony; Yoo, James J.

In: Methods, Vol. 47, No. 2, 01.02.2009, p. 98-103.

Research output: Contribution to journalArticle

Eberli, Daniel ; Soker, Shay ; Atala, Anthony ; Yoo, James J. / Optimization of human skeletal muscle precursor cell culture and myofiber formation in vitro. In: Methods. 2009 ; Vol. 47, No. 2. pp. 98-103.
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