Heat‐inactivated coelomic fluid of the earthworm <i>Perionyx excavatus</i> is a possible alternative source for fetal bovine serum in animal cell culture

Vasantha, Niranjan Chellathurai; Rajagopalan, Kamarajan; Christyraj, Jackson Durairaj Selvan; Chelladurai, Karthikeyan Subbiahanadar; Ganesan, Mijithra; Azhagesan, Ananthaselvam; Pushpabai, Rajesh Rajaian; Mohan, Manikandan; Christyraj, Johnson Retnaraj Samuel Selvan

doi:10.1002/btpr.2817

Cited by 9 publications

(6 citation statements)

References 15 publications

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“…Nevertheless, in the last six decades researchers have been trying to alternate FBS with different sources include hormones [ 24 ], pituitary extracts, chick embryo extracts, bovine milk fractions, bovine colostrum or platelet lysates [ 14 ], bovine ocular fluid [ 25 ], fish serum [ 26 ], plant origin [ 27 ] and sericin protein [ 28 ], other protein fractions from plant extracts, called vegetal serum [ 29 ] and earthworm coelomic fluid [ 30 ]. However, the serum-free chemically defined media is the good one for clinical and pharmaceutical productions since it is specific to every cell type [ 31 ].…”

Section: Efforts Made To Reduce or Replace Fbsmentioning

confidence: 99%

“…But in the case of the adherent cell, the presence of fibrinolytic enzymes in CF does not allow the cells to attach and grow [ 78 ]. Nonetheless, the crude CF has a toxic effect on cells and interestingly our group [ 30 ] recently encounter and evaluated the feasibility of using 10% heat-inactivated coelomic fluid (HI-CF) from the earthworm, P. excavatus , for culturing mouse primary fibroblast cells and HeLa cell lines without FBS. The cells supplemented with HI-CF were grown well and show a similar growth rate and better cellular viability when compared to FBS.…”

Section: Efforts Made To Reduce or Replace Fbsmentioning

confidence: 99%

“…But if we can combine HPL- (HI-CF) with the ratio 1:9 or 2:8 and 10% supplemented it in Basal media which will be enough for the cells to attach and proliferate (A) which will be similar to FBS media (B). Detailed methodology for the preparation of HPL discussed by Burnouf, T et al 2016 and HI-CF [ 30 ]. …”

Section: Future Perspectivesmentioning

confidence: 99%

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Alternative to FBS in animal cell culture - An overview and future perspective

Chelladurai

Christyraj

Rajagopalan

et al. 2021

Heliyon

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Fetal bovine serum (FBS) is a widely used growth supplement in the in vitro culturing of animal and human cells, tissues and organs, notably due to the occurrence of abundant micro-and macronutrients, along with growth factors. Over the years, increasing demand, high price, batch-to-batch variability in quality and composition, increasing ethical concerns lead to the search for an alternative to FBS. Several approaches have been suggested and employed in the past, but none is implemented as widely as FBS, and each supplement has its own disadvantages. In this review, we described the importance of FBS in cell culture, discussed the issues associated with FBS use and presented the efforts made in the recent past to reduce or replace FBS. The potential of four different alternative sources to FBS, namely, bovine ocular fluid, sericin protein, human platelet lysate and earthworm heat inactivated coelomic fluid was evaluated. In the end, we present the conceptual perspective using the Human Platelet Lysate (HPL) and earthworm Heat Inactivated Coelomic Fluid (HI-CF) combination to alternate FBS and its context in scientific and economic impacts.

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Section: Efforts Made To Reduce or Replace Fbsmentioning

confidence: 99%

Section: Efforts Made To Reduce or Replace Fbsmentioning

confidence: 99%

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Alternative to FBS in animal cell culture - An overview and future perspective

Chelladurai

Christyraj

Rajagopalan

et al. 2021

Heliyon

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“…The coelomic fluid (CF) of the earthworm Perionyx excavatus was proven to be a potential alternative for FBS in animal culture. However, it lacks cell attachment due to the presence of fibrinolytic enzymes 8,9 . In this study, we have formulated a heat inactivated coelomic fluid (HI-CF) containing serum free media (f-HICF), which aids in the growth, attachment, and proliferation of the adherent cells.…”

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confidence: 99%

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

Rossan Mathews,

Subramaniam,

Venkatachalam

et al. 2024

Sci Rep

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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.

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“…In order to collect the ECF, electric shock method was conducted (MohamedJaabir et al 2011;Chellathurai Vasantha et al 2019). Earthworms suspended in the media containing Hank's balanced salt solution (HBSS) and phosphate buffered saline (PBS) (1:8, w/v, pH 7.0), and then were subjected to electric shock (3V, 30 sec, 1 min interval for recovery and repeated 5 times).…”

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confidence: 99%

Biomarker Responses in Earthworms Lumbricus Terrestris L. (Annelida: Lumbricidae) to Artificial Soils Contaminated with Dimethyl Phthalate

Farsani¹,

Arabi²,

Shadkhast³

2021

Preprint

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In ecotoxicological studies, oxidative damages and antioxidant defense responses in earthworms have been used as hallmarks for assessing the environmental pollution. Phthalates, which are esters of phthalic acid (PAEs) are primarily used to enhance plasticity of industrial polymers, and are an unavoidable part of modern life. In the present study, ecotoxicological effects of dimethyl phthalate (DMP) were investigated in the earthworms Lumbricus terrestris L., at different concentrations (0, 0.1, 1, 10 and 50 mg kg-1) on the 7th and 14th days of exposure in artificial soils. We evaluated the effects of DMP on oxidative stress (OS) biomarkers including malondialdehyde (MDA/LPO) level and the activities of antioxidant enzymes catalase (CAT) and glutathione S-transferase (GST), and carbonylated proteins (CP) content in earthworm’s whole body extract. The neutral red retention (NRR)-time of coelomocytes and total antioxidant capacity (TAC) were also measured in the earthworm’s coelomic fluid (ECF). Upon exposure to DMP, OS status was induced in a time and concentration-dependent manner. The MDA level, CAT and GST activities, and CP content were found to be increased, but the TAC value and NRR-times were decreased in all DMP-treated groups on 7th and 14th days of exposure. The toxicity of DMP was generally greater in the groups with 14 day toxicant exposure. Briefly, our resultes defined that DMP is a source of oxidative damages in the earthworms. However, for detailed elucidation of the mechanisms associated with DMP ecotoxicity and its adverse impacts in the soli organisms, further unraveling is needed.

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Heat‐inactivated coelomic fluid of the earthworm Perionyx excavatus is a possible alternative source for fetal bovine serum in animal cell culture

Cited by 9 publications

References 15 publications

Alternative to FBS in animal cell culture - An overview and future perspective

Alternative to FBS in animal cell culture - An overview and future perspective

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

Biomarker Responses in Earthworms Lumbricus Terrestris L. (Annelida: Lumbricidae) to Artificial Soils Contaminated with Dimethyl Phthalate

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