One-Step Purification of Human Skeletal Muscle Myoblasts and Subsequent Expansion Using Laminin-Coated Surface

Shiplu Roy Chowdhury, Annis Binti Ismail, Sia Chye Chee, Mohd Suffian Bin Laupa, Fadhlun Binti Jaffri, Salfarina Ezrina Mohmad Saberi, Ruszymah Idrus

Research output: Contribution to journalArticle

6 Citations (Scopus)

Abstract

Skeletal myoblasts have been extensively used to study muscle growth and differentiation, and were recently tested for their application as cell therapy and as a gene delivery system to treat muscle and nonmuscle diseases. However, contamination of fibroblasts in isolated cells from skeletal muscle is one of the long-standing problems for routine expansion. This study aimed to establish a simple one-step process to purify myoblasts and maintain their purity during expansion. Mixed cells were preplated serially on laminin- and collagen type I-coated surfaces in a different array for 5, 10, and 15 min. Immunocytochemical staining with antibodies specific to myoblasts was performed to evaluate myoblast attachment efficiency, purity, and yield. It was found that laminin-coated surface favors the attachment of myoblasts. Highest myoblast purity of 78.9%±6.8% was achieved by 5 min of preplating only on the laminin-coated surface with a yield of 56.9%±3.3%. Primary cells, isolated from skeletal muscle (n=4), confirm the enhancement of purity through preplating on laminin-coated surface for 5 min. Subsequent expansion after preplating enhanced myoblast purity due to an increase in myoblast growth than fibroblasts. Myoblast purity of ∼98% was achieved when another preplating was performed during passaging. In conclusion, myoblasts can be purified and efficiently expanded in one step by preplating on laminin-coated surface, which is a simple and robust technique.

Original languageEnglish
Pages (from-to)1135-1142
Number of pages8
JournalTissue Engineering - Part C: Methods
Volume21
Issue number11
DOIs
Publication statusPublished - 1 Nov 2015

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Skeletal Myoblasts
Myoblasts
Laminin
Purification
Muscle
Skeletal Muscle
Fibroblasts
Collagen Type I
Collagen
Antibodies
Contamination
Genes
Gene Transfer Techniques
Muscles
Growth
Cell- and Tissue-Based Therapy
Staining and Labeling

ASJC Scopus subject areas

  • Biomedical Engineering
  • Bioengineering
  • Medicine (miscellaneous)

Cite this

One-Step Purification of Human Skeletal Muscle Myoblasts and Subsequent Expansion Using Laminin-Coated Surface. / Chowdhury, Shiplu Roy; Ismail, Annis Binti; Chee, Sia Chye; Laupa, Mohd Suffian Bin; Jaffri, Fadhlun Binti; Mohmad Saberi, Salfarina Ezrina; Idrus, Ruszymah.

In: Tissue Engineering - Part C: Methods, Vol. 21, No. 11, 01.11.2015, p. 1135-1142.

Research output: Contribution to journalArticle

Chowdhury, Shiplu Roy ; Ismail, Annis Binti ; Chee, Sia Chye ; Laupa, Mohd Suffian Bin ; Jaffri, Fadhlun Binti ; Mohmad Saberi, Salfarina Ezrina ; Idrus, Ruszymah. / One-Step Purification of Human Skeletal Muscle Myoblasts and Subsequent Expansion Using Laminin-Coated Surface. In: Tissue Engineering - Part C: Methods. 2015 ; Vol. 21, No. 11. pp. 1135-1142.
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