Pig Pluripotent Stem Cells as a Candidate for Biomedical Application

Choi, Kwang‐Hwan; Lee, Chang Kyu

doi:10.12750/jarb.34.3.139

Cited by 17 publications

(19 citation statements)

References 52 publications

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“…Muscle stem cells are generally cultured as proliferating myoblasts because the satellite cells cannot be maintained in vitro without losing their properties ( Gilbert et al, 2010 ; Montarras et al, 2005 ). The in vitro culture of stem cells is accomplished by mimicking the in vivo cellular niche ( Choi and Lee, 2019 ). To trigger proliferation in vitro , the isolated satellite cells are cultured with various cytokines and signaling molecules, which are secreted from the surrounding cells, such as stromal and interstitial cells, and muscle stem cells themselves ( Yin et al, 2013 ).…”

Section: Introductionmentioning

confidence: 99%

Optimization of Culture Conditions for Maintaining Pig Muscle Stem Cells In Vitro

Choi¹,

Yoon²,

Kim³

et al. 2020

Food Sci Anim Resour

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Muscle stem cells isolated from domestic animals, including cows and pigs, were recently spotlighted as candidates for the production of alternative protein resources, so-called cultured meat or lab-grown meat. In the present study, we aimed to optimize the in vitro culture conditions for the long-term expansion of pig muscle stem cells via the screening of various signaling molecules. Pig muscle stem cells were collected from the biceps femoris muscles of 3-d-old crossbred pigs (Landrace×Yorkshire×Duroc, LYD) and cultured in minimum essential medium-based growth media. However, the pig muscle stem cells gradually lost their proliferation ability and featured morphologies during the long-term culture over two weeks. To find suitable in vitro culture conditions for an extended period, skeletal muscle growth medium-2, including epidermal growth factor (EGF), dexamethasone, and a p38 inhibitor (SB203580), was used to support the stemness of the pig muscle stem cells. Interestingly, pig muscle stem cells were stably maintained in a long-term culture without loss of the expression of myogenic marker genes as determined by PCR analysis. Immunostaining analysis showed that the stem cells were capable of myogenic differentiation after multiple passaging. Therefore, we found that basal culture conditions containing EGF, dexamethasone, and a p38 inhibitor were suitable for maintaining pig muscle stem cells during expanded culture in vitro . This culture method may be applied for the production of cultured meat and further basic research on muscle development in the pig.

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Section: Introductionmentioning

confidence: 99%

Optimization of Culture Conditions for Maintaining Pig Muscle Stem Cells In Vitro

Choi¹,

Yoon²,

Kim³

et al. 2020

Food Sci Anim Resour

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“…Given the role of pig as a signi cant livestock for supplying meat, the myogenic reprogramming of pig cells can be applied to increase agricultural yield and produce cultured meat. Additionally, pigs provide biomedical applications in preclinical studies for human disease because of their anatomical and physiological similarities with humans (52,53). For example, transdifferentiated pig muscle cells can offer a cell source for skeletal muscle disease modeling and drug screening in regenerative medicine.…”

Section: Discussionmentioning

confidence: 99%

Myogenic Transdifferentiation of Pig Fibroblasts Using the Porcine MYOD1 Inducible Vector

Jeong¹,

Choi²,

Kim³

et al. 2020

Preprint

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Background Myogenic transdifferentiation can be accomplished through ectopic MYOD1 expression, which is facilitated by various signaling pathways associated with myogenesis. In this study, we attempted to transdifferentiate pig embryonic fibroblasts (PEFs) myogenically into skeletal muscle through overexpression of the pig MYOD1 gene and modulation of the FGF, TGF-β, WNT, and cAMP signaling pathways.Results The MYOD1 overexpression vector was constructed based on comparative sequence analysis, demonstrating that pig MYOD1 has evolutionarily conserved domains across various species. Although forced MYOD1 expression through these vectors triggered the expression of endogenous muscle markers, transdifferentiated muscle cells from fibroblasts were not observed. Therefore, various signaling molecules, including FGF2, SB431542, CHIR99021, and forskolin, along with MYOD1 overexpression were applied to enhance the myogenic reprogramming. The modified conditions led to the derivation of myotubes and activation of muscle markers in PEFs, as determined by qPCR and immunostaining. Notably, a sarcomere-like structure was observed, indicating that terminally differentiated skeletal muscle could be obtained from transdifferentiated cells.Conclusions In summary, we established a protocol for reprogramming MYOD1-overexpressing PEFs into the mature skeletal muscle using signaling molecules. Our myogenic reprogramming can be used as a cell source for muscle disease models in regenerative medicine and the production of cultured meat in cellular agriculture.

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“…Accordingly, research into pig PSCs such as embryonic stem cells (ESCs) and induced pluripotent stem cells (iPSCs) is required for understanding their comparative developmental biology and medical/agricultural applications. However, to date, no stable and well-characterized pig PSCs have been derived due to a lack of understanding of pig pluripotency networks and suitable culture conditions [3,4]. Instead, ES-like cells, having no in vivo differentiation ability, and transgene-dependent partially reprogrammed iPSCs have been derived and researched over the last few decades [5].…”

Section: Objectivementioning

confidence: 99%

Transcriptome profiling of pluripotent pig embryonic stem cells originating from uni- and biparental embryos

Choi

Lee

et al. 2020

BMC Res Notes

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Objectives: Pig pluripotent stem cells have tremendous potential because the pig is a valuable animal as both an agricultural resource and as a preclinical model of human therapy. To date, a lack of understanding of pig pluripotency has inhibited the derivation of embryonic stem cells (ESCs) and transgene-free induced pluripotent stem cells. Therefore, there has been no accessible or reliable transcriptome data for researching the genuine pig pluripotency network. Our previous study isolated authentic pig ESCs, which had teratoma-forming and direct differentiation ability, that were derived by activating the FGF2, ACTIVIN A, and WNT pathways. Here, we aimed to provide detailed information on transcriptome data of the newly derived pig ESCs and perform a comparative analysis with pig preimplantation embryo transcriptomes in a public database. Data description: The transcriptome data of ESCs derived from in vitro fertilized and parthenogenetic embryos were generated by HiSeq 2500. Then, differentially expressed genes (DEGs) from each sample were compared with fibroblasts, and gene expression profiling was carried out for comparative analysis. Our data, as the first transcriptome dataset for genuine pig pluripotent cells, could be a general reference for explaining the molecular mechanism of species-specific pluripotency and improving understanding of the embryo development of domestic animals.

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Pig Pluripotent Stem Cells as a Candidate for Biomedical Application

Cited by 17 publications

References 52 publications

Optimization of Culture Conditions for Maintaining Pig Muscle Stem Cells In Vitro

Optimization of Culture Conditions for Maintaining Pig Muscle Stem Cells In Vitro

Myogenic Transdifferentiation of Pig Fibroblasts Using the Porcine MYOD1 Inducible Vector

Transcriptome profiling of pluripotent pig embryonic stem cells originating from uni- and biparental embryos

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