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

Effect of standard and physiological cell culture temperatures on in vitro proliferation and differentiation of primary broiler chicken pectoralis major muscle satellite cells

“…T10282). Cells were diluted in proliferation media, as described in Flees et al (2022) , consisting of low-glucose DMEM with 10% chicken serum (Millipore Sigma, Burlington, MA, Cat. C5405) + 5% horse serum + 1% antibiotic/antimycotic + 0.1% gentamicin to achieve 40,000 live cells per well in triplicate on 24-well, gelatin-coated, tissue culture plates.…”

Current Research, Industrialization Status, and Future Perspective of Cultured Meat

“…In the culture of MSCs, the basal medium and serum concentrations are known to be closely related to the cell proliferation rate and myotube formation ( McFarland et al, 1988 ). In a broiler MSC culture experiment based on culture medium composition, DMEM was found to be more effective than McCoy’s 5A medium in terms of proliferation rate and MRF expression ( Flees et al, 2022 ). In addition, the common view that a low glucose content is effective for chicken and bovine MSC proliferation when using DMEM as basal medium was confirmed ( Flees et al, 2022 ; Zygmunt et al, 2023 ).…”