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objective:To study systematically the abilities of skeletal muscle cells in synthesizing,processing and secreting recombinant proteins in vitro.Methods:Primary murine myoblasts from SCID mice (SCID-MB) and C2C12,a muse myoblast cell line,were trans fected with muscle specific (pdLMe4bAh ff m) or non-specif ic (LIXSN) vectors encoding human factor IX (FIX). The capacities of the transgenic cells in producing re combinant FIX were compared in the levels of intracellular and secreted FIX protein as well as FIX mRNA in primary muscle cells and C2C12 cells. Results:Both primary muscle cells and C2C12 cells can efficiently trans late and process FIX protein. Myotubes derived from the SCID-MB produced 1 800~2 000 ng FIX/106 cells/ 24 h in culture with specific activity of 95% ~ 103%. C2C12 cells can synthesize and process recombinant FIX as efficiently as primary muscle cells,but their secretion ability is less efficient in comparison with primary cells. Conclusion:This observation indicates that primary cells should be used in gene therapy related research projects and care should be taken while explaining the results derived from established cell lines.
Objective: To study systematically the abilities of skeletal muscle cells in synthesizing, processing and secreting recombinant proteins in vitro. Methods: Primary murine myoblasts from SCID mice (SCID-MB) and C2C12, a muse myoblast cell line, were trans fected with muscle specific (pdLMe4bAh ff m) or non-specif ic (LIXSN) vectors encoding human factor IX (FIX). The capacities of the transgenic cells in producing re combinant FIX were compared in the levels of intracellular and secreted FIX protein as well as FIX mRNA in Primary muscle cells and C2C12 cells. Results: Both primary muscle cells and C2C12 cells efficiently efficiently late and process FIX protein. Myotubes derived from the SCID-MB produced 1 800-2 000 ng FIX / 106 cells / 24 h in culture with specific activity of 95% ~ 103%. C2C12 cells can synthesize and process recombinant FIX as efficiently as primary muscle cells, but their secretion ability is less efficient in comparison with primary cells. that primary cells should be used in gene therapy related research projects and care should be taken while explaining the results derived from established cell lines.