Biochemical and functional characterization of heat-inactivated coelomic fluid from earthworms as a potential alternative for fetal bovine serum in animal cell culture

• Published in: Scientific reports 2024-03, Vol.14 (1), p.5606-5606, Article 5606
• Main Authors: Rossan Mathews, Melinda Grace, Subramaniam, Ravichandran, Venkatachalam, Saravanakumar, Selvan Christyraj, Johnson Retnaraj Samuel, Yesudhason, Beryl Vedha, Kalimuthu, Kalishwaralal, Mohan, Manikandan, Selvan Christyraj, Jackson Durairaj
• Format: Article
• Language: English
• Subjects: 631/1647; 631/45; 631/61; Adherent cells; Animals; Cell culture; Cell Culture Techniques - methods; Cell lines; Cell proliferation; Cells, Cultured; Coelomic fluid; Contamination; Culture Media - chemistry; Culture Media, Serum-Free; Earthworm; Endotoxins; Ethics; Fetal bovine serum; Fetal calf serum; HeLa Cells; Hot Temperature; Humanities and Social Sciences; Humans; Lipids; Micronutrients; multidisciplinary; Nutrient availability; Oligochaeta; Perionyx excavatus; Science; Science (multidisciplinary); Serum Albumin, Bovine; Serum free media; Sterility; Trace elements; Trace elements (nutrients); Vitamins
• ISSN: 2045-2322; 2045-2322
• DOI: 10.1038/s41598-024-56169-0

Fetal bovine serum (FBS) plays a pivotal role in animal cell culture. Due to ethical and scientific issues, searching for an alternative, comprising the three R’s (Refinement, Reduction and Replacement) gained global attention. In this context, we have identified the heat inactivated coelomic fluid (HI-CF) of the earthworm, Perionyx excavatus as a potential alternative for FBS. Briefly, we formulated HI-CF (f-HICF) containing serum free medium which can aid the growth, attachment, and proliferation of adherent cells, similar to FBS. In this study, we investigated the biochemical characterization, sterility, stability, formulation, and functional analysis of HI-CF as a supplement in culturing animal cells. Notably, vitamins, micronutrients, proteins, lipids, and trace elements are identified and compared with FBS for effective normalization of the serum free media. HI-CF is tested to be devoid of endotoxin and mycoplasma contamination thus can qualify the cell culture grade. The f-HICF serum free media was prepared, optimised, and tested with A549, HeLa, 3T3, Vero and C2C12 cell lines. Our results conclude that f-HICF is a potential alternative to FBS, in accordance with ethical concern; compliance with 3R's; lack of unintended antibody interactions; presence of macro and micronutrients; simple extraction; cost-effectiveness and availability.