Cell line development starts with cell isolation, disinfection of the cell source, mechanical, or enzymatic tissue dissociation, cell growth in media, cell characterization, passaging, and developing immortalized cell lines.
Bovine satellite cells, chicken cells, porcine cells, and fish cells were isolated for propagation, to assess initial doubling times and survival with passaging; subsets of these cells will be further characterized for immortalization
Primary cultures of oyster muscle cells were generated from Crassostrea virginica, an economically important oyster specie in Eastern US that currently has no stable cell line available for cultivated meat production; combinations of minimal L15 media and 5% fetal bovine serum was optimal
The team has moved through the assessment of various disinfectants, media combinations, and plate coatings to obtain conditions to reduce contamination during primary isolations, which is a particular challenge for marine species
Studies are continuing on immortalization and subculture of the various cell lines.
Directed differentiation of zebrafish embryonic stem cells (ZEM2S) into myogenic cell lines is underway, with the testing of growth factor cocktails and serum levels to monitor myogenic differentiation
Several species including blue crab, cray fish, oyster, clam, Pompano, American Eel, and Flounder are under investigation
