Myogenic Cell Lines Derived from Transgenic Mice Carrying a Thermolabile T Antigen: A Model System for the Derivation of Tissue-Specific and Mutation-Specific Cell Lines

Novel murine clonal cell lines either express slow or mixed (fast and slow) muscle markers following differentiation in vitro

“…Cultures of all these clones easily lead to expression of adult MHC isoforms, in contrast to other classically used rodent myogenic cell lines (C2, L6, BCH3, etc.). The satellite cells from the immortomouse have also been shown to contribute to muscle fiber regeneration in vivo (Morgan et al, 1994;Cousins et al, 2004). WTt cells, at day10 of differentiation, were cultured with 10% fetal calf serum (FCS) and 2% chicken embryo extract (CEE), and 10 g of cell protein extracts were deposited per well.…”

“…They have also been shown to form normal muscle, following implantation into mdx mice, and not tumors (Morgan et al, 1994), unlike other classic myogenic cell lines such as C2 (Morgan et al, 2002). They continue to form normal muscle after serial passage, indicating that they retain at least some properties of stem cells (Morgan et al, 1994). The clones derived from these transgenic mice differentiate into a strikingly more mature phenotype than all other widely used myoblast clones or primary cultures of satellite cells (LaFramboise et al, 2003;Miller et al, 2003;Morgan et al, 2002).…”

“…Original myogenic cell lines were derived from limb satellite cells of neonatal H-2K b -tsA58 transgenic mice as described (Morgan et al, 1994). In addition, clones were derived from EDL, LD, or Soleus muscles of adult mice, by dissecting out the muscles first before isolating the satellite cells.…”

“…These cell lines grow with the ease of spontaneously transformed cell lines and retain full capacity for differentiation. They have also been shown to form normal muscle, following implantation into mdx mice, and not tumors (Morgan et al, 1994), unlike other classic myogenic cell lines such as C2 (Morgan et al, 2002). They continue to form normal muscle after serial passage, indicating that they retain at least some properties of stem cells (Morgan et al, 1994).…”

“…We took advantage of the properties of cells derived from the H-2K b -tsA58 transgenic mouse, also called immortomouse (Jat et al, 1991;Morgan et al, 1994), in which cells are conditionally immortalized by a temperature sensitive T-antigen under control of a gamma-interferon inducible promoter. This enabled us to isolate clones derived from single satellite cells from neonatal muscle, or adult Soleus, Extensor digitorum longus, or Longissimus dorsi muscles, expand them in culture and investigate their phenotypes upon differentiation into myotubes.…”