title: One-Step Purification of Human Skeletal Muscle Myoblasts and Subsequent Expansion Using Laminin-Coated Surface.
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.
authors.affiliation: 1 Tissue Engineering Centre, Faculty of Medicine, Universiti Kebangsaan Malaysia Medical Center , Universiti Kebangsaan Malaysia, Kuala Lumpur, Malaysia .
authors.forename: Shiplu Roy
authors.initials: SR
authors.lastname: Chowdhury
authors.affiliation: 1 Tissue Engineering Centre, Faculty of Medicine, Universiti Kebangsaan Malaysia Medical Center , Universiti Kebangsaan Malaysia, Kuala Lumpur, Malaysia .
authors.forename: Annis
authors.initials: A
authors.lastname: binti Ismail
authors.affiliation: 1 Tissue Engineering Centre, Faculty of Medicine, Universiti Kebangsaan Malaysia Medical Center , Universiti Kebangsaan Malaysia, Kuala Lumpur, Malaysia .
authors.forename: Sia Chye
authors.initials: SC
authors.lastname: Chee
authors.affiliation: 1 Tissue Engineering Centre, Faculty of Medicine, Universiti Kebangsaan Malaysia Medical Center , Universiti Kebangsaan Malaysia, Kuala Lumpur, Malaysia .
authors.forename: Mohd Suffian
authors.initials: MS
authors.lastname: bin Laupa
authors.affiliation: 1 Tissue Engineering Centre, Faculty of Medicine, Universiti Kebangsaan Malaysia Medical Center , Universiti Kebangsaan Malaysia, Kuala Lumpur, Malaysia .
authors.forename: Fadhlun
authors.initials: F
authors.lastname: binti Jaffri
authors.affiliation: 1 Tissue Engineering Centre, Faculty of Medicine, Universiti Kebangsaan Malaysia Medical Center , Universiti Kebangsaan Malaysia, Kuala Lumpur, Malaysia .
authors.forename: Salfarina Ezrina Mohmad
authors.initials: SE
authors.lastname: Saberi
authors.affiliation: 1 Tissue Engineering Centre, Faculty of Medicine, Universiti Kebangsaan Malaysia Medical Center , Universiti Kebangsaan Malaysia, Kuala Lumpur, Malaysia .|2 Department of physiology, Faculty of Medicine, Universiti Kebangsaan Malaysia , Kuala Lumpur, Malaysia .
authors.forename: Ruszymah Bt Hj
authors.initials: RB
authors.lastname: Idrus
mesh_terms: D000818:Animals; D002448:Cell Adhesion / Q000187:drug effects; D049109:Cell Proliferation / Q000187:drug effects; D002469:Cell Separation / Q000379:methods*; D003094:Collagen / Q000494:pharmacology; D005347:Fibroblasts / Q000166:cytology / Q000187:drug effects; D006801:Humans; D007797:Laminin / Q000494:pharmacology*; D018482:Muscle, Skeletal / Q000166:cytology*; D032446:Myoblasts / Q000166:cytology* / Q000187:drug effects; D051381:Rats
chemical_list: D007797:Laminin; D003094:Collagen