Fibroblast growth factor, epidermal growth factor, insulin‐like growth factors, and platelet‐derived growth factor‐BB stimulate proliferation of clonally derived porcine myogenic satellite cells

Abstract

Clonally derived cultures of porcine skeletal muscle satellite cells were developed. The mitogenic effects of basic fibroblast growth factor (bFGF), epidermal growth factor (EGF), insulin-like growth factors (IGF-I and -II), and platelet-derived growth factors (PDGF-AA and -BB) were examined. Individually, bFGF, IGFs, and PDGF-BB stimulated proliferation of porcine satellite cells grown in basal serum-free medium or Minimum Essential Medium containing 2% fetal bovine serum (MEM-2% FBS). EGF stimulated proliferation in MEM-2% FBS, but neither EGF nor PDGF-AA were mitogenic when added to serum-free medium. The interactions among bFGF, EGF, IGF-I, and PDGF-BB were examined in serum-free medium, using growth factor concentrations shown in dose-response experiments to induce maximal proliferative responses (10 ng/ml bFGF, EGF and PDGF-BB, and 50 ng/ml IGF-I). The combination of bFGF and IGF-I dramatically increased proliferation, and IGF-I also synergized with EGF to increase proliferation. EGF, IGF-I, and bFGF interacted with PDGF-BB to stimulate proliferation. With the exception of EGF and bFGF, combinations of two growth factors typically resulted in greater than additive responses. Simultaneous exposure of satellite cells to bFGF, PDGF-BB, EGF, and IGF-I produced a fivefold increase in DNA compared to cells grown in basal serum-free medium. Elimination of EGF did not reduce the mitogenic response, yet removal of IGF-I, bFGF, or PDGF-BB reduced proliferation by approximately 40, 20, and 10%, respectively. These mitogens are likely physiological regulators of porcine satellite cell activity.

A Requirement for Fibroblast Growth Factor in Regulation of Skeletal Muscle Growth and Differentiation Cannot Be Replaced by Activation of Platelet-Derived Growth Factor Signaling Pathways

“…Therefore, FGF may function intracellularly in addition to activating its cell surface receptor, and this intracellular function may be critical for FGF-dependent cell proliferation or repression of differentiation. Although PDGF does not affect MM14 cell differentiation, it has been shown to repress myogenesis and promote proliferation of other muscle cell lines (25,62) and primary muscle cell cultures (14,62). In these studies, the mechanisms involved were not investigated, and it is possible that PDGF was stimulating FGF activity indirectly.…”

Prospectus of cultured meat—advancing meat alternatives

“…Serum-free media reduce operating costs and process variability while lessening the potential source of infectious agents (Froud 1999). Improvements in the composition of commercially available cell culture media have enhanced our ability to successfully culture many types of animal cells and serum-free media have been developed to support in vitro myosatellite cell cultures from the turkey (McFarland et al 1991), sheep (Dodson and Mathison 1988) and pig (Doumit et al 1993). Variations among different serum-free media outline the fact that satellite cells from different species have different requirements and respond differentially to certain additives (Dodson et al 1996).…”

“…Furthermore, formulation may be required to change over the course of the culturing process from proliferation period to the differentiation and maturation period, requiring different set of factors. A multitude of regulatory factors have been identified as being capable of inducing myosatellite cell proliferation (Cheng et al 2006), and the regulation of meat animalderived myosatellite cells by hormones, polypeptide growth factors and extracellular matrix proteins has also been investigated (Dodson et al 1996;Doumit et al 1993). Purified growth factors or hormones may be supplemented into the media from an external source such as transgenic bacterial, plant or animal species which produce recombinant proteins (Houdebine 2009).…”

Associated analysis of single nucleotide polymorphisms found on exon 3 of the IGF-1 gene with Tibetan miniature pig growth traits

“…The IGF-1 gene promotes muscle cell differentiation in the muscle cell formation process (Hembree et al, 1991;Buckingham et al, 2003;O'Sullivan et al, 2011). The IGF-1 gene also regulates the growth of skeletal muscle in the process of skeletal muscle growth (Doumit et al, 1993;Lamberson et al, 1995;Davis et al, 2002;Schlegel et al, 2010;Wang et al, 2011Wang et al, , 2012. In addition, the IGF-1 gene stimulates myoblast terminal differentiation by inducing the expression of the myogenin (MyoG) gene (Florini et al, 1991;Yano et al, 1999).…”