A repurposing screen of 29 media mixtures derived from 15 commercial serum-free media products identifies potential cost reductions for industrial animal cell cultivation

Abstract We performed a repurposing screen of 15 commercial serum-free products, across 29 different media mix options, in Nunc™ EasYFlask™ cell culture flasks/CELLBIND® 6-well plates, agnostic of their recommended application or cell type. 20% v/v FBS DMEM was replaced with a given serum-free mix in a single step, and growth monitored over 6 days for the mouse muscle cell line, Sol8. 20 serum-free media mixes reduced or failed to increase growth, while 9 increased cell growth. 'Mix D', Fishers base medium containing 2% v/v Ultroser G serum alternative, showed the best combination of cell number increase and volumetric media cost reduction: 33% lower than 20% v/v FBS DMEM. Further Nunc™ EasYFlask™ cell culture flasks/CELLBIND® 6-well plates experiments showed that Mix D supported levels of Sol8 cell growth comparable to 20% v/v FBS DMEM, but with lower levels of metabolic activity. Comparable growth in Mix D was observed for Sol8 cells that had undergone a period of adaptation to the serum-free medium, and clonally-derived Sol8 sub-populations. Mix D also matched serum-containing media performance in supporting growth of the porcine muscle cell line, pMYOSC, the porcine pre-adipocyte fat cell line, pADSC and the mouse pre-adipocyte cell line, 3T3 L1 in Nunc™ EasYFlask™ cell culture flasks/CELLBIND® 6-well plates. 3T3 L1 cells cultivated in Mix D retained their capacity to differentiate into adipocytes, a cell type important for animal fat cultivation. The observation that Mix D supported growth of four different animal cell types at levels comparable to serum-containing medium recommends this formulation, and this repurposing screen approach, to researchers seeking serum alternatives.