An effective cytokine combination for ex vivo expansion of porcine muscle stem cells

Lei, Qingzi; Li, Mei; Du, Guocheng; Zhou, Jingwen; Guan, Xin

doi:10.1016/j.fbio.2022.101571

Cited by 19 publications

(14 citation statements)

References 42 publications

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“…Cells were further purified by fluorescence-activated cell sorting with the CD31 − /CD45 − /CD56 + /CD29 + cell surface marker combination. 31…”

Section: Methodsmentioning

confidence: 99%

“…Cells were further purified by fluorescence-activated cell sorting with the CD31 − /CD45 − /CD56 + /CD29 + cell surface marker combination. 31 Purified pMuSCs were seeded on Matrigel (Corning, NY, USA)-coated 24 well plates and cultured with 1 mL growth medium (GM, consisted of DMEM with 10% FBS and 1% P/S) at 37°C in a humidified 5% CO 2 incubator. The medium was changed every two days and cells were passaged using 0.25% trypsin digestion when reaching 60% confluency.…”

Section: Isolation and Culture Of Pmuscsmentioning

confidence: 99%

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Vitamin C enhances the ex vivo proliferation of porcine muscle stem cells for cultured meat production

Fang

Zhang

et al. 2022

Food Funct.

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“…Cells were further purified by fluorescence-activated cell sorting with the CD31 − /CD45 − /CD56 + /CD29 + cell surface marker combination. 31…”

Section: Methodsmentioning

confidence: 99%

Section: Isolation and Culture Of Pmuscsmentioning

confidence: 99%

Vitamin C enhances the ex vivo proliferation of porcine muscle stem cells for cultured meat production

Fang

Zhang

et al. 2022

Food Funct.

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“…The basal differentiation medium (DM) was DMEM plus 2% heat-inactivated HS and 5 mg/mL P/S. Primary porcine satellite cells were isolated from large white pigs as reported previously and cultured on collagen-coated plates [ 27 ]. The culture medium and procedures for satellite cells were the same as C2C12 cells.…”

Section: Methodsmentioning

confidence: 99%

“…Cells were permeabilized with 0.5% Triton X-100 and blocked with 1% BSA for 1 h, then incubated with anti-MHC primary antibody at 4 °C overnight and subsequently with CoraLite488-conjugated secondary antibodies (Proteintech, Wuhan, China) at room temperature for 1 h. Images were captured with a fluorescence microscope (MF52-M, Mshot, Guangzhou, China) and the MHC-positive myotubes were calculated to quantify the differentiation efficiency. The fusion index was determined by dividing the number of nuclei in MHC-positive cells and by total number of nuclei in each image [ 27 ]. In some experiments that analyze mitophagy, cells were infected with an adenovirus encoding GFP-LC3 (Beyotime, Shagnhai, China) to visualize autophagosomes 48 h prior to permeabilization and the anti-TOMM20 antibody was used to label mitochondria.…”

Section: Methodsmentioning

confidence: 99%

IGF-1 Signaling Regulates Mitochondrial Remodeling during Myogenic Differentiation

et al. 2022

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Skeletal muscle is essential for locomotion, metabolism, and protein homeostasis in the body. Mitochondria have been considered as a key target to regulate metabolic switch during myo-genesis. The insulin-like growth factor 1 (IGF-1) signaling through the AKT/mammalian target of rapamycin (mTOR) pathway has a well-documented role in promoting muscle growth and regeneration, but whether it is involved in mitochondrial behavior and function remains un-examined. In this study, we investigated the effect of IGF-1 signaling on mitochondrial remodeling during myogenic differentiation. The results demonstrated that IGF-1 signaling stimulated mitochondrial biogenesis by increasing mitochondrial DNA copy number and expression of genes such as Cox7a1, Tfb1m, and Ppargc1a. Moreover, the level of mitophagy in differentiating myoblasts elevated significantly with IGF-1 treatment, which contributed to mitochondrial turnover. Peroxisome proliferator-activated receptor gamma coactivator 1-alpha (PGC-1α) and BCL2/adenovirus E1B 19 kDa protein-interacting protein 3 (BNIP3) were identified as two key mediators of IGF-1-induced mitochondrial biogenesis and mitophagy, respectively. In addition, IGF-1 supplementation could alleviate impaired myoblast differentiation caused by mitophagy deficiency, as evidenced by increased fusion index and myosin heavy chain expression. These findings provide new insights into the role of IGF-1 signaling and suggest that IGF-1 signaling can serve as a target for the research and development of drugs and nutrients that support muscle growth and regeneration.

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“…The proliferation and activation of SCs differentiation can be regulated by extracellular signaling molecules (e.g., growth factors-GFs, cytokines, and myokines), which have different biological effects on skeletal muscle function and myogenesis via various signaling pathways, such as Ras/MAPK, JAK/STAT and PI3K/Akt [194]. In this regard, these molecules may have potential roles in CM production.…”

Section: Biochemical Cuesmentioning

confidence: 99%

Using Vertebrate Stem and Progenitor Cells for Cellular Agriculture, State-of-the-Art, Challenges, and Future Perspectives

et al. 2022

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Global food systems are under significant pressure to provide enough food, particularly protein-rich foods whose demand is on the rise in times of crisis and inflation, as presently existing due to post-COVID-19 pandemic effects and ongoing conflict in Ukraine and resulting in looming food insecurity, according to FAO. Cultivated meat (CM) and cultivated seafood (CS) are protein-rich alternatives for traditional meat and fish that are obtained via cellular agriculture (CA) i.e., tissue engineering for food applications. Stem and progenitor cells are the building blocks and starting point for any CA bioprocess. This review presents CA-relevant vertebrate cell types and procedures needed for their myogenic and adipogenic differentiation since muscle and fat tissue are the primary target tissues for CM/CS production. The review also describes existing challenges, such as a need for immortalized cell lines, or physical and biochemical parameters needed for enhanced meat/fat culture efficiency and ways to address them.

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An effective cytokine combination for ex vivo expansion of porcine muscle stem cells

Cited by 19 publications

References 42 publications

Vitamin C enhances the ex vivo proliferation of porcine muscle stem cells for cultured meat production

Vitamin C enhances the ex vivo proliferation of porcine muscle stem cells for cultured meat production

IGF-1 Signaling Regulates Mitochondrial Remodeling during Myogenic Differentiation

Using Vertebrate Stem and Progenitor Cells for Cellular Agriculture, State-of-the-Art, Challenges, and Future Perspectives

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