The global demand for animal proteins is projected to rise by 14% by 2030, amplifying the environmental toll of conventional animal-based protein production. Cultivated meat technology can alleviate the growing demand for protein and address the environmental and ethical concerns associated with conventional livestock farming. However, it faces a critical challenge: the high cost of cell culture media, primarily due to the use of Fetal Bovine Serum (FBS). Substituting serum with protein hydrolysates reduces the production expense of cultivated meat products and promotes establishing a sustainable food system. This study explores black soldier fly larvae (Hermetia illucens) as an emerging ethical and cost-effective alternative protein source to replace serum in media, particularly for cultivated meat production. The development of BSFL protein isolate involved defatting the larva, followed by protein extraction. The protein isolate was then hydrolyzed using an enzyme to produce BSFL hydrolysates. The goal was to supplement the protein isolate and hydrolysates with a serum-free media (B8) and determine their efficacy in replacing the 20% serum requirement for the cell culture of Bovine Satellite Cells. The BSFL protein isolate developed had a crude protein content of 80.42% and an amino acid composition conducive to cell proliferation. Experimental concentrations, ranging from 0.006 mg/ml for hydrolysate to 0.06 mg/ml for protein isolate, exhibited enhanced cell growth. Data from dsDNA quantification revealed no significant difference in growth between cells fed serum-containing growth media (BSC-GM) and BSFL protein hydrolysate (BSFLH_1h) over a short-term study. Results from the multi-passage growth study revealed that BSFLH_1h significantly improved cell growth compared to B8 over 4 passages. However, its doubling time was slower than BSC-GM. Additionally, it was observed that the protein isolate and hydrolysate were cytotoxic at higher concentrations. In the future, identifying and removing the cytotoxic compounds can further optimize the media composition. Immunostaining using Pax7 and DAPI identified supplemented media-maintained satellite cell identity of Bovine satellite cells, offering crucial insights into cellular proliferation. Furthermore, since each cell type requires varying serum and nutrients, testing these isolates and hydrolysates on different cell lines can provide better insight into creating a universal serum-free media. / Master of Science in Life Sciences / The global demand for animal proteins is projected to rise by 14% by 2030, amplifying the environmental toll of conventional animal-based protein production. Meat, dairy, aquaculture, and eggs significantly contribute to food-related emissions and occupy a vast portion of global farmland. Cultivated meat production can alleviate the growing demand for protein and address the environmental and ethical concerns associated with conventional livestock farming. Currently, the production of cultivated meat faces a significant hurdle: the high cost of culture media, primarily attributed to the use of Fetal Bovine Serum (FBS).
Substituting serum with protein isolates or hydrolysates reduce the production expense of cultivated meat products and promotes a sustainable food system. Protein isolate and hydrolysates derived from black soldier fly larvae (Hermetia illucens) are rich in protein and essential amino acids and can be used as a cost-effective alternative to serum in cell culture media. The protein isolate and hydrolysates derived from BSFL were tested as supplements to a serum-free media (B8) to evaluate their effectiveness in supporting the growth of Bovine Satellite Cells. The protein hydrolysate demonstrated enhanced cell growth at experimental concentrations. However, it could not completely replace serum requirements without slowing cell growth. Despite challenges such as cytotoxicity at higher concentrations, our study suggests that further refinements and application on various cell types can assist in creating a sustainable and affordable serum-free media for cultivated meat production.
| Identifer | oai:union.ndltd.org:VTETD/oai:vtechworks.lib.vt.edu:10919/117301 |
| Date | 03 January 2024 |
| Creators | Garg, Palak |
| Contributors | Food Science and Technology, Ovissipour, Reza, Zarei, Mohammad, Strawn, Laura K. |
| Publisher | Virginia Tech |
| Source Sets | Virginia Tech Theses and Dissertation |
| Language | English |
| Detected Language | English |
| Type | Thesis |
| Format | ETD, application/pdf, application/pdf |
| Rights | In Copyright, http://rightsstatements.org/vocab/InC/1.0/ |
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