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A serum-free media formulation for cultured meat production supports bovine satellite cell differentiation in the absence of serum starvation

ABSTRACT: This SuperSeries is composed of the SubSeries listed below.

ORGANISM(S): Bos taurus

PROVIDER: GSE173199 | GEO | 2021/10/22

REPOSITORIES: GEO

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A serum-free media formulation for cultured meat production supports bovine satellite cell differentiation in the absence of serum starvation [SF_Diff]

Project description:Production of cultured meat requires the robust differentiation of satellite cells into mature muscle fibers without the use of animal-derived components. Current protocols induce myogenic differentiation in vitro through serum starvation, an abrupt reduction in serum concentration. Here, we used RNA sequencing to investigate the transcriptomic remodelling of bovine satellite cells during myogenic differentiation induced by serum starvation. We characterized canonical myogenic gene expression, and identified surface receptors upregulated during the early phase of differentiation. Supplementation of ligands to these receptors enabled the formulation of a chemically defined media that induced differentiation in the absence of serum starvation and/or transgene expression. Serum-free myogenic differentiation was of similar extent to that induced by serum starvation, as evaluated by transcriptome analysis, protein expression and the presence of a functional contractile apparatus. Moreover, the serum-free differentiation media supported the fabrication of mature three-dimensional bioartificial muscle constructs, demonstrating its suitability for cultured beef production.

2021-10-22 | GSE173196 | GEO

A serum-free media formulation for cultured meat production supports bovine satellite cell differentiation in the absence of serum starvation [Timecourse]

2021-10-22 | GSE173198 | GEO

A serum-free media formulation for cultured meat production supports bovine satellite cell differentiation in the absence of serum starvation

Project description:A serum-free media formulation for cultured meat production supports bovine satellite cell differentiation in the absence of serum starvation

| PRJNA724671 | ENA

A serum-free media formulation for cultured meat production supports bovine satellite cell differentiation in the absence of serum starvation [SF_Diff]

Project description:A serum-free media formulation for cultured meat production supports bovine satellite cell differentiation in the absence of serum starvation [SF_Diff]

| PRJNA724677 | ENA

A serum-free media formulation for cultured meat production supports bovine satellite cell differentiation in the absence of serum starvation [Timecourse]

Project description:A serum-free media formulation for cultured meat production supports bovine satellite cell differentiation in the absence of serum starvation [Timecourse]

| PRJNA724678 | ENA

Extracellular small non-coding RNA contaminants in fetal bovine serum and serum-free media.

Project description:In the research field of extracellular vesicles (EVs), the use of fetal bovine serum (FBS) depleted of EVs for in vitro studies is advocated to eliminate the confounding effects of media derived EVs. EV-depleted FBS may either be prepared by ultracentrifugation or purchased commercially. Nevertheless, these preparations do not guarantee an RNA-free FBS for in vitro use. In this study we address the RNA contamination issue, of small non-coding (nc)RNA in vesicular or non-vesicular fractions of FBS, ultracentrifugation EV-depleted FBS, commercial EV-depleted FBS, and in our recently developed filtration based EV-depleted FBS. Commercially available serum- and xeno-free defined media were also screened for small ncRNA contamination. Our small ncRNA sequencing data showed that all EV-depleted media and commercially available defined media contained small ncRNA contaminants. Out of the different FBS preparations studied, our ultrafiltration-based method for EV depletion performed the best in depleting miRNAs. Certain miRNAs such miR-122 and miR-203a proved difficult to remove completely and were found in all media. Compared to miRNAs, other small ncRNA (snRNA, Y RNA, snoRNA, and piRNA) were difficult to eliminate from all the studied media. Additionally, our tested defined media contained miRNAs and other small ncRNAs, albeit at a much lower level than in serum preparations. Our study showed that no media is free of small ncRNA contaminants. Therefore, in order to screen for baseline RNA contamination in culturing media, RNA sequencing data should be carefully controlled by adding a media sample as a control. This should be a mandatory step before performing cell culture experiments in order to eliminate the confounding effects of media.

| S-EPMC6445286 | biostudies-literature

Questioning the purity of the media - Extracellular small non-coding RNA contaminants in fetal bovine serum and serum-free media

Project description:In the research field of extracellular vesicles (EVs), the use of EV-depleted fetal bovine serum (FBS) for in vitro studies is highly recommended to eliminate the confounding effects of media derived EVs. EV-depleted FBS may either be prepared by ultracentrifugation or bought commercially, nevertheless these depletion methods do not guarantee an RNA-free preparation. In this study we have addressed the RNA contamination issue in FBS, ultracentrifuged EV-depleted FBS, commercially available EV-depleted FBS, and also from our recently developed filtration based EV depleted FBS. Commercially available serum-free, xeno-free defined media were also screened for RNA contamination.

2019-04-30 | GSE120594 | GEO

A serum-free medium formulation efficiently supports isolation and propagation of canine adipose-derived mesenchymal stem/stromal cells.

Project description:Medium containing Fetal Bovine Serum (FBS) provides a supportive environment for isolation and expansion of mesenchymal stromal/stem cells (MSCs); however, the inherent variability of FBS may contribute to inconsistencies in cell growth and yield between batches of stem cell products. For this reason, we set out to develop a serum-free medium capable of supporting the in vitro expansion of MSCs. First a naïve serum-free medium was formulated by Sato's approach. Once it was established that the naïve serum-free medium supported the expansion of canine adipose-derived MSCs (Ad-MSCs), the serum-free medium was optimized by addition of growth factors. Combinations of growth factors were chosen and compared by their effect on cell proliferation and colony formation. Growth characteristics of canine adipose-derived MSCs cultured in the serum-free medium were comparable to those cultured in standard FBS containing medium. In addition, cell surface marker expression and differentiation potential of serum-free and FBS-based cultures were also comparable. However, a commercial serum-free medium developed for human MSC culture did not support growth of canine Ad-MSCs. In summary, canine Ad-MSCs isolated and cultured in serum-free medium retained the basic characteristics of MSCs cultured in FBS containing medium.

| S-EPMC6392232 | biostudies-literature

Revealing suspension adapation of CHO cell in serum free media

Project description:Purpose : We performed strand-specific RNA-seq analysis of CHO cells according to adaptation trajectory to identify the specific genes responded to the suspension adaptation. Result : Cell growth, replication, and nucleotide metabolism categories were enriched in up regulated clusters whereas cellular community category which contains focal adhesion related genes were enriched in down regulated clusters

2015-11-19 | GSE75094 | GEO

Comparative analysis of mesenchymal stem cells cultivated in serum free media

Project description:Differential expression analysis of mRNAs and proteins showed that the expression of genes related with apoptosis, immune response, and inflammatory response were significantly up-regulated in ADSCs cultivated in FBS containing media.

2022-04-05 | GSE200086 | GEO

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