Expression of muscle‐specific isozymes of phosphorylase and creatine kinase in human muscle fibers cultured aneurally in serum‐free, hormonally/chemically enriched medium

Abstract

Primary cultures of muscle cells derived from a biopsied adult human skeletal muscle were grown up to 6 weeks in a hormonally/chemically enriched serum-free medium. The expression of muscle-specific isozymes of creatine kinase, glycogen phosphorylase, and phosphoglycerate mutase, indicative of muscle cell maturation, was studied after 1, 4 and 6 weeks of growth. The maturation of muscle fibers cultured in serum-free medium was comparable to that achieved by muscle fibers cultured in medium containing 10% serum and supplemented with growth factors (insulin, epidermal growth factor, and fibroblastic growth factor) and was greater than that achieved in medium containing 10% serum only. Our study demonstrates that adult human muscle can be cultured aneurally for a long period of time in a serum-free medium, and that it can achieve a high degree of maturation. This study provides an important basis for investigations related to: (1) assessment of the influence of individual components of the medium on human muscle maturation in culture; (2) studies of regulation of abnormal gene expression in diseased human muscle cultured in serum-fre medium.

Activation of the Unfolded Protein Response in Sporadic Inclusion-Body Myositis but Not in HereditaryGNEInclusion-Body Myopathy

“…Primary cultures of normal human muscle were established as routinely performed in our laboratory (37–39), from archived satellite cells of portions of diagnostic muscle biopsies from: a) 3 h-IBM patients carrying M712T and 1 patient carrying N519S GNE mutation, and b) 5 different patients who, after all tests had been performed, were considered free of muscle disease. Each experiment was established from satellite cells derived from a different muscle biopsy but not all experiments were performed on each sample.…”

Cystatin C colocalizes with amyloid‐β and coimmunoprecipitates with amyloid‐β precursor protein in sporadic inclusion‐body myositis muscles

“…1990). This medium promotes differentiation of aneurally cultured human muscle fibers and enables their maintenance for several weeks (Pegolo et al . 1990).…”

“…Tissue cultures of normal human muscle were established, as we described, from satellite cells of portions of diagnostic muscle biopsies from patients who, after all tests were performed, were considered free of muscle disease (reviewed in . Cultures were initially maintained in F14 medium (Gibco/ Invitrogen, Carlsbad, CA, USA) supplemented with 10% fetal bovine serum (Hyclone, Logan, UT, USA), insulin (10 lg/mL, Sigma, St Louis, MO, USA), and an antibiotic/antimycotic mixture (Sigma), and subsequently maintained in the serum-free medium (Pegolo et al 1990). This medium promotes differentiation of aneurally cultured human muscle fibers and enables their maintenance for several weeks (Pegolo et al 1990).…”

“…Cultures were initially maintained in F14 medium (Gibco/ Invitrogen, Carlsbad, CA, USA) supplemented with 10% fetal bovine serum (Hyclone, Logan, UT, USA), insulin (10 lg/mL, Sigma, St Louis, MO, USA), and an antibiotic/antimycotic mixture (Sigma), and subsequently maintained in the serum-free medium (Pegolo et al 1990). This medium promotes differentiation of aneurally cultured human muscle fibers and enables their maintenance for several weeks (Pegolo et al 1990). Cultures were processed for immunocytochemistry and immunoblots as described (Askanas et al 1996b(Askanas et al , 1997.…”

Hydrocortisone influences voltage‐dependent L‐type calcium channels in cultured human skeletal muscle

“…When the serum-free medium was used, cells were first grown for up to 10 days in the "aneural medium," and then, at the time when HC treatment was initiated, switched to the serum-free medium (Pegolo et al, 1990).…”