Isolation, expansion and differentiation of myogenic progenitors:
Laminin-521 is a superior substrate for both short-term and long-term myogenic cell culture
During myogenesis, laminin-521 and laminin-511 are the main isoforms surrounding myogenic progenitors (Gawlik & Durbeej, 2011). Indeed, recent data presented by researchers at Icagen Inc. demonstrate that the laminin-521 (LN-521) cell culture matrix maintains the differentiation potential of mouse and human satellite cell-derived myoblasts, even during long-term culture expansion (Penton et al., 2016). LN-521 support increased proliferation during expansion and superior differentiation with myotube hypertrophy, larger myotubes and higher amounts of nuclei per myotube. Moreover, Penton et al. show that LN-521 support more consistent and reliable differentiation over long-term culture and is the only substrate facilitating high-level fusion following long-term culture. LN-521 support increased differentiation potential without altering the traditional Pax7/MyoD paradigm and the results are translational across several mouse backgrounds, human cells and disease states. Counterintuitively, culture of satellite cell-derived myoblasts on laminin-211, the native laminin isoform in resting skeletal muscle, result in low proliferation and poor differentiation (Penton et al., 2016).
Laminin-211 is the most abundant laminin isoform in adult skeletal muscle
Laminin-211 is the most abundant laminin isoform in the basement membrane of adult skeletal muscle with integrin α7β1 as its major receptor (Holmberg & Durbeej, 2013). Laminin is crucial for normal muscle function, evident from naturally occurring mutations in laminin genes. Mutations in the gene encoding laminin a2 chain are the most common cause of congenital muscular dystrophy type 1A. For a review on laminin-211 in skeletal muscle function, see Holmberg & Durbeej, 2013.
Laminin-421 and laminin-111 are also expressed in adult skeletal muscle in small amounts, preferably at junctional regions (Patton et al., 1999)
Laminin β-2 immunohistochemical staining of
human skeletal muscle show strong membranous
immunoreactivity in myocytes. Anti-LAMB2 antibody
AMAb91096 (Atlas Antibodies).
- Laminin-211 in skeletal muscle function. Holmberg & Durbeej, Cell Adh Migr., 2013
- Skeletal muscle laminin and MDC1A: pathogenesis and treatment strategies. Gawlik & Durbeej. Skelet Muscle, 2011
- Laminin 521 maintains differentiation potential of mouse and human satellite cell-derived myoblasts during long-term culture expansion. Penton et al. Skeletal Muscle, 2016
- Distribution of ten laminin chains in dystrophic and regenerating muscles. Patton et al. Neuromuscul Disord., 1999