Volume 4 | Issue 1 | January-June 2018 | Page: 16-22 | David Livingstone, Albert A Kota1, Sanjay K Chilbule, Karthikeyan Rajagopal, Sukria Nayak, Vrisha Madhuri
Authors: David Livingstone, Albert A Kota , Sanjay K Chilbule, Karthikeyan Rajagopal, Sukria Nayak , Vrisha Madhuri
Department of Orthopaedics, Paediatric Orthopaedics Unit, 1Department of Surgery, Unit IV, Christian Medical College, Vellore, Tamil Nadu, India
Address of Correspondence
Dr. Vrisha Madhuri,
Paediatric Orthopaedics Unit, Christian Medical College, Vellore – 632 009, Tamil Nadu, India.
Introduction: Satellite cells are a resident population of stem cells beneath the basal lamina of mature skeletal muscle fibers. Their capacity to regenerate muscle makes them a potentially ideal source for human cell therapy with respect to muscle-related diseases such as urinary and fecal incontinence, and others. In this study, we describe a protocol to isolate, expand in-vitro, and characterize human muscle satellite cells from the rectus abdominis muscle. Materials and Methods: Muscle biopsies from human donors were harvested, digested using collagenase type II, and then plated on extracellular matrix-coated plates.
Results: Immunocytochemistry revealed that satellite cells on day 8 were 70–80% Pax7 positive; in contrast, cells expanded until day 12 showed 50–75% positivity for Pax7. The real-time polymerase chain reaction for day 8 culture indicated four-fold increase in Pax3 and Pax7 gene expression, four-fold increase in MyoD gene expression, and five-fold increase in Myf5 gene expression.
Conclusion: These findings suggest that satellite cells can be cultured until day 8 for translational purposes. The protocol described here is modest, operational, and reproducible and involves only basic cell culture equipment.
Keywords: Cell therapy, Human skeletal muscle, Myoblast, Satellite cells, Sphincter injuries, Tissue regeneration
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|How to Cite this Article: Livingstone D, Kota AA, Chilbule SK, Rajagopal K, Nayak S, Madhuri V | Isolation, in-vitro expansion, and characterization of human muscle satellite cells from the rectus abdominis muscle | January-June 2018; 4(1): 16-22.