Biology subjects

Ovissipour, R.

Publications and source records attributed to Ovissipour, R..

7 recordsLinked to original sources

bioRxiv2025

Integrated Multi-Omics Characterization of CDK4/hTERT-Immortalized Ovine Satellite Cells for Cultivated Meat Applications

Cultivated meat production requires stable, scalable cell lines that maintain essential biological characteristics. Here, we establish and comprehensively characterize CDK4/hTERT immortalized ovine satellite cells using flow cytometry-based isolation and integrated transcriptomic-proteomic analysis. Flow cytometry identified highly purified satellite cells which were successfully immortalized and maintained stable proliferation (14-22h doubling time) through 67 passages. Multi-omics analysis revealed progressive molecular evolution with 754-1180 differentially expressed genes and 960 differentially expressed proteins, demonstrating coordinated transcriptional and post-transcriptional adaptations. Immortalization triggered species-specific extracellular matrix remodeling and inflammatory activation while preserving essential myogenic markers and apoptotic machinery. Immunofluorescence analysis demonstrated that key proteins, including PAX7, remained detectable despite transcriptional downregulation and absence of corresponding signals in mass spectrometry. Pathway analysis confirmed non-oncogenic profiles with enhanced metabolic flexibility and preserved differentiation capacity. These findings provide the first comprehensive molecular characterization of immortalized ovine satellite cells, revealing species-specific responses crucial for cultivated meat applications and establishing a framework for systematic cell line safety assessment.

bioengineering↗

bioRxiv2024

Evaluating the Efficacy of Serum-Free Media Supplemented with Protein Isolates for Bovine Satellite Cell Proliferation: A Sustainable Approach for Cultivated Meat Production

This study investigates the short- and long-term efficacy of serum-free media supplemented with protein isolates from algae, alfalfa, silkworm pupae, and grasshoppers for the proliferation of bovine satellite cells (BSCs). Fresh and spent media were analyzed to monitor metabolites, while cell proliferation was assessed using the CyQUANT assay. The results of the short-term growth study indicated that a lower concentration of alfalfa protein isolate (0.05 mg/mL) significantly enhanced cell proliferation, achieving a 1.47-fold increase compared to basal media, thereby demonstrating its potential as a viable alternative to fetal bovine serum (FBS). The capability of the developed serum-free media for cell passaging was examined using different coating strategies, with vitronectin outperforming others; however, it was unable to support the long-term growth of cells compared to FBS-containing media. Given the critical role of glutamine, this study also evaluates the impact of L-glutamine supplementation at different concentrations on cell growth and metabolism in alfalfa-based media. However, while glutamine supplementation showed trends toward increased cell growth, the enhancements were not statistically significant. Based on the results, this study highlights the potential of alfalfa protein isolate as a promising component of serum-free media for BSC proliferation and underscores the need for continued research into alternative protein sources and media formulations to support the sustainable and ethical production of cultivated meat.

bioengineering↗

bioRxiv2024

Assessment of Plant- and Microbial-Derived Protein Hydrolysates as Sustainable for Fetal Bovine Serum in Seafood Cell Culture Media

This study seeks to explore alternatives by substituting or reducing the conventional 10% serum concentration in Zebrafish embryonic stem cell (ESC) growth media with protein hydrolysates sourced from peas, mushrooms, yeast, and algae. Notably, algae exhibited the highest protein content, optimal amino acid balance, and favorable functional properties. When applied at concentrations ranging from 1 to 10 mg/mL, all protein hydrolysates demonstrated pro-apoptotic effects and inhibited cell growth, particularly when used in conjunction with 10% serum. However, concentrations ranging from 0.001 to 0.1 mg/mL displayed anti-apoptotic properties and promoted cell proliferation. The study found that media containing 1% or 2.5% serum, along with 0.01 mg/mL of protein hydrolysates, supported cell growth effectively. Lactate Dehydrogenase (LDH) Activity served as an indicator of cell health and integrity under the specified conditions of protein hydrolysate supplementation. Cells cultured in serum-free media exhibited significantly decreased cell membrane integrity (P < 0.05) compared to those in regular media or media containing low serum (1% and 2.5%) along with low concentrations (0.01 mg/mL) of protein hydrolysates. Furthermore, analysis of Greenhouse Gas emissions (GHG) suggested that media formulations containing 1% serum combined with low concentrations of protein hydrolysates present a sustainable approach for cell-based seafood production.

bioengineering↗

bioRxiv2024

Black Soldier Fly Bioconversion to Cultivated Meat Media Components Using Blue Catfish Gut Microbiome

Developing low-cost media is one of the major challenges in the cellular agriculture domain. Thus, this study aimed to develop low-cost media for cell-cultivated seafood using gut-microbial community-assisted fermentation. Black soldier fly larvae (Hermetia illucens) were used as the substrate and exposed to gut microbial communities isolated from Blue catfish (Ictalurus furcatus). In the first step, BSFL slurry was subjected to enzymatic digestion, using pepsin and trypsin to mimic animal digestive processes. The results showed a 2.8% degree of hydrolysis after digestion with pepsin and an additional 5.9% after digestion with trypsin. In the second step, two fermentation approaches were tested, including the direct addition of gut homogenate to the hydrolysates (fermentation A) and the establishment of microbial cultures from the gut homogenate before fermentation (fermentation B). Both fermentations resulted in similar protein content and degree of hydrolysis. Fermentation led to a decrease in species richness, with the loss of important chitinase and protease-producing genera such as Pseudomonas and Clostridiaceae. However, there was an increase in Paraclostridium and members of the Enterobacteriaceae. In addition, the effect of fermented hydrolysates from BSFL on the proliferation of zebrafish embryo fibroblasts was tested in comparison to fetal bovine serum (FBS) in in vitro cell cultivation. Lower concentrations of FBS resulted in decreased cell density and altered cell morphology. The supplementation of hydrolysate B at high peptide concentrations had cytotoxic effects on the cells, while at lower peptide concentrations, it improved cell proliferation only in cultures with 2.5% FBS.

bioengineering↗

bioRxiv2023

Decellularized Banana Leaves: Eco-Friendly Scaffolds for Cell-Based Seafood

Cellular agriculture, as an emerging food production system, holds potential to address sustainability, food security, and agricultural resilience. Within the cell-based meat supply chain, one of the key steps is scaffolding. In this study, we assessed decellularized banana leaves, various coating materials, and different cell seeding strategies to determine their effects on cell viability, cell growth, cell alignment, and the response of the materials to thermal processing. The efficiency of decellularization was verified through DNA quantification, which decreased from 445 ng/mg in fresh banana leaves to non-detectable levels in the decellularized samples. This was further confirmed by FTIR and PCA modeling. Cell viability exceeded 98% on uncoated, soy-coated, and gelatin-coated samples of the decellularized banana leaves. Alignment of cells on gelatin-coated samples was the highest among the samples, with a dominant orientation of 65.8{degrees}, compared to soy-coated and uncoated samples with dominant orientations of 9.2{degrees} and -6.3{degrees}, respectively. In terms of quality attributes, the kinetics of shrinkage indicated that coating with soy and the presence of cells increased the activation energy due to the higher energy required for protein denaturation. Moreover, the kinetics of area changes in plain scaffolds without cells followed a first-order pattern, while with seeded cells a second-order pattern was followed. In summary, decellularized banana leaves present a sustainable and suitable biomaterial to support cells towards future needs related to meat production.

synthetic biology↗

bioRxiv2023

Investigating the Impact of Microplastics on Fish Muscle Cell Proliferation and Differentiation: Enhancing Food Safety in Cultivated Meat Production

Cultivated meat, a sustainable alternative to traditional livestock farming, has gained attention for its potential environmental and health benefits. However, concerns about microplastic contamination pose challenges, especially when sourcing cells from marine organisms prone to microplastic bioaccumulation. Additionally, the pervasive presence of microplastics in laboratory settings, ingredients, and during the production, increases the risk of unintentional contamination. This study focused on Atlantic mackerel (Scomber scombrus) skeletal muscle cell lines to examine the effects of microplastic exposure, represented by fluorescent polyethylene microspheres (10-45 {micro}m) on cell performance including cell proliferation, cell viability, gene expression, and differentiation processes critical for cultivated meat production. The results revealed significant impacts on cell attachment and proliferation at microplastic concentrations of 1 {micro}g/mL, 10 {micro}g/mL, and 50 {micro}g/mL. Notably, the 10 {micro}g/mL concentration exerted the most pronounced effects on cell viability during both attachment and proliferation phases. While the results indicated that both microplastic concentration and size influence cell viability, cell differentiation remained unaffected, and additional contributing factors require further investigation. These findings underscore the necessity of thoroughly exploring microplastic-cell interactions to ensure food safety and safeguard health within the burgeoning cultivated meat industry.

systems biology↗

bioRxiv2022

Evaluating the potential of marine invertebrate and insect protein hydrolysates to reduce fetal bovine serum in cell culture media for cultivated fish production

The use of fetal bovine serum (FBS) and the price of the cell culture media are the key constraints for developing serum-free cost-effective media. This study aims to replace or reduce the typical 10% serum application in fish cell culture media by applying protein hydrolysates from insects and marine invertebrate species for the growth of Zebrafish embryonic stem cells (ESC) as the model organism. Protein hydrolysates were produced from Black soldier fly (BSF), cricket, oyster, mussel, and lugworm with high protein content, suitable functional properties, adequate amino acids composition, and the degree of hydrolysis from 18.24 to 33.52%. Protein hydrolysates at low concentrations from 0.001 to 0.1 mg/mL in combination with 1 and 2.5% serum significantly increased cell growth compared to the control groups (5 and 10% serum) (P < 0.05). All protein hydrolysates with concentrations of 1 and 10 mg/mL were found to be toxic to cells and significantly reduced cell growth and performance (P < 0.05). However, except for cricket, all hydrolysates were able to restore or significantly increase cell growth and viability with 50% less serum at a concentration of 0.001, 0.01, and 0.1 mg/mL. Although cell growth was enhanced at lower concentrations of protein hydrolysates, cell morphology was altered due to the lack of serum. Lactate dehydrogenase (LDH) activity results indicated that BSF and lugworm hydrolysates did not alter the cell membrane. In addition, light and fluorescence imaging revealed that cell morphological features were comparable to the 10% serum control group. Overall, lugworm and BSF hydrolysates reduced serum by up to 90% while preserving excellent cell health.

systems biology↗