Effect of Different Basal Culture Media and Sera Type Combinations on Primary Broiler Chicken Muscle Satellite Cell Heterogeneity during Proliferation and Differentiation

SIMPLE SUMMARY: Little consistency in the literature exists for optimal culture conditions for proliferating and differentiating primary broiler chicken muscle satellite cells regarding basal culture media, proliferation sera, and differentiation sera. This experiment assessed primary satellite cell proliferation and differentiation when cultured in different combinations of basal media and sera. Cells were cultured in different basal media: low glucose Dulbecco’s Modified Eagle’s medium, McCoy’s 5A, and high glucose Dulbecco’s Modified Eagle’s medium. Each media was supplemented with 15% chicken serum, or a combination of 5% horse serum + 10% chicken serum during proliferation while 3% horse serum or 3% chicken serum were supplemented during differentiation. Cultures were immunofluorescence stained for myogenic regulatory factors at different time points during proliferation and differentiation. During proliferation and differentiation, cells cultured in Dulbecco’s Modified Eagle’s medium tended to have higher proportions of myogenic cells expressing myogenic regulatory factors and promoted satellite cell fusion into myotubes compared with McCoy’s 5A. Low glucose media, glucose concentration similar to circulating glucose concentrations in broilers, combined with sera published in the literature may be the optimal culture media to promote satellite cell proliferation and differentiation. ABSTRACT: The objective of this experiment was to access primary satellite cell (SC) proliferation and differentiation when cultured in different combinations of basal media and sera due to little consistency being published on the optimal culture media for primary broiler chicken satellite cells. Cells were cultured in one of three different basal media: McCoy’s 5A, high glucose Dulbecco’s Modified Eagle’s medium (DMEM), and low glucose DMEM. Media were supplemented with 15% chicken serum (CS) or a combination of 5% horse serum (HS) + 10% CS during proliferation while 3% HS or 3% CS were added to the media during differentiation. Cultures were immunofluorescence stained for myogenic regulatory factors (MRF) at 48, 72, and 96 h post-plating for proliferation (Pax7, MyoD, and Myf-5) and 96 h post-proliferation during differentiation (Pax7 and MyoD), including MF20 to assess fusion. Cells cultured in Dulbecco’s Modified Eagle’s medium tended to have higher proportions of myogenic cells expressing MRF during proliferation and promoted fusion into myotubes compared with McCoy’s 5A during differentiation. Culturing primary SC in low glucose media, glucose concentrations similar to circulating glucose concentrations in broilers, HSCS during proliferation and CS during differentiation, appears to be optimal for promoting broiler chicken satellite cell proliferation and differentiation.