Generation of embryonic stem‐like cells from in vivo‐derived porcine blastocysts at a low concentration of basic fibroblast growth factor

Hj, Cha; Ji, Yun; Han, Ning; Hy, Kim; Baek, Song Ee; Sh, Lee; J, Lee; Lee, Eric; Ck, Park; Lee, S-T.

doi:10.1111/rda.13088

Cited by 11 publications

(6 citation statements)

References 40 publications

(47 reference statements)

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“…At present, each of the porcine embryonic stem cell‐like cells (pESLCs) exhibited some features of pluripotency; however, these cells cannot maintain self‐renewal for a long time 5‐7 . Further investigation showed that only individual cell lines exhibited inefficient chimeras 8 .…”

Section: Introductionmentioning

confidence: 99%

BCL2 enhances survival of porcine pluripotent stem cells through promoting FGFR2

et al. 2020

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Objectives The establishment of porcine pluripotent stem cells (pPSCs) is still a critical topic. However, all pPSCs were failed to contribute to efficient chimeric pig and were extremely sensitive to changes of culture conditions. This study aimed to investigate the role of BCL2 in pPSCs and further explain the mechanism. Materials and Methods Porcine BCL2 gene was cloned and overexpressed in porcine induce pluripotent stem cells (piPSCs). Digital RNA‐seq was performed to explain the mechanism of anti‐apoptosis. Finally, the cells carrying BCL2 were injected into mouse early embryo to evaluate its chimeric ability. Results Here, we found that overexpression of porcine BCL2 gene significantly improved the survivability of piPSCs and the efficiency of embryonic chimerism, and did not wreck the pluripotency of piPSCs. Furthermore, the Digital RNA‐seq analysis revealed that BCL2, as a downstream gene of the PI3K signal pathway, enhanced the expression of PI3K signal pathway receptors, such as FGFR2, and further promoted oxidoreductases activity and lipid metabolism, thus maintaining the survival and pluripotency of piPSCs. Conclusion Our data not only suggested that porcine BCL2 gene could enhance the survivability and chimeric ability of pPSCs, but also explained the positive feedback mechanism in this process, providing strong support for the chimeric experiment of pPSCs.

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Section: Introductionmentioning

confidence: 99%

BCL2 enhances survival of porcine pluripotent stem cells through promoting FGFR2

et al. 2020

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“…In this study, fibroblasts were induced from ADSCs. [ 7 8 ] In studies on the effects of bFGF to promote fibroblast functions, bFGF concentrations vary from 0.005 to 2000 ng/ml,[ 2 9 10 11 12 ] with no defined optimal concentration. EGF is a widely used growth factor to promote the repair of skin fibroblasts.…”

Section: Introductionmentioning

confidence: 99%

Effect of Optimized Concentrations of Basic Fibroblast Growth Factor and Epidermal Growth Factor on Proliferation of Fibroblasts and Expression of Collagen

Jia

Zhou

Chang

et al. 2018

Chinese Medical Journal

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Background:Fibroblasts were the main seed cells in the studies of tissue engineering of the pelvic floor ligament. Basic fibroblast growth factor (bFGF) and epidermal growth factor (EGF) were widely studied but at various concentrations. This study aimed to optimize the concentrations of combined bFGF and EGF by evaluating their effects on proliferation and collagen secretion of fibroblasts.Methods:Fibroblasts were differentiated from rat adipose mesenchymal stem cells (ADSCs). Flow cytometry and immunohistochemistry were used for cell identification. The growth factors were applied at concentrations of 0, 1, 10, and 100 ng/ml as three groups: (1) bFGF alone, (2) EGF alone, and (3) bFGF mixed with EGF. Cell proliferation was evaluated by Cell Counting Kit-8 assays. Expression of Type I and III collagen (Col-I and Col-III) mRNAs was evaluated by real-time quantitative reverse transcription-polymerase chain reaction. Statistical analysis was performed with SPSS software and GraphPad Prism using one-way analysis of variance and multiple t-test.Results:ADSCs were successfully isolated from rat adipose tissue as identified by expression of typical surface markers CD29, CD44, CD90, and CD45 in flow cytometry. Fibroblasts induced from ADSC, compared with ADSCs, were with higher mRNA expression levels of Col I and Col III (F = 1.29, P = 0.0390). bFGF, EGF, and the mixture of bFGF with EGF can enhanced fibroblasts proliferation, and the concentration of 10 ng/ml of the mixture of bFGF with EGF displayed most effectively (all P < 0.05). The expression levels of Col-I and Col-III mRNAs in fibroblasts displayed significant increases in the 10 ng/ml bFGF combined with EGF group (all P < 0.05).Conclusions:The optimal concentration of both bFGF and EGF to promote cell proliferation and collagen expression in fibroblasts was 10 ng/ml at which fibroblasts grew faster and secreted more Type I and III collagens into the extracellular matrix, which might contribute to the stability of the pelvic floor microenvironment.

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“…Therefore, most of piPSCs also fail to contribute to chimeras with exceptions that are only detected by genomic PCR analysis 39,40 . Porcine embryonic stem cell‐like cells (pESLCs) exhibited some features of pluripotency, but these cells could not maintain self‐renewal for a long time 58‐61 . The generation of pESCs and acquisition of imperfect piPSCs always fail because the porcine endogenous pluripotent network cannot be maintained stably.…”

Section: Discussionmentioning

confidence: 99%

Histone demethylase complexes KDM3A and KDM3B cooperate with OCT4/SOX2 to define a pluripotency gene regulatory network

Zhu¹,

Wu²,

et al. 2021

The FASEB Journal

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The pluripotency gene regulatory network of porcine induced pluripotent stem cells(piPSCs), especially in epigenetics, remains elusive. To determine the biological function of epigenetics, we cultured piPSCs in different culture conditions. We found that activation of pluripotent gene-and pluripotency-related pathways requires the erasure of H3K9 methylation modification which was further influenced by mouse embryonic fibroblast (MEF) served feeder. By dissecting the dynamic change of H3K9 methylation during loss of pluripotency, we demonstrated that the H3K9 demethylases KDM3A and KDM3B regulated global H3K9me2/me3 level and that their co-depletion led to the collapse of the pluripotency gene regulatory network.Immunoprecipitation-mass spectrometry (IP-MS) provided evidence that KDM3A and KDM3B formed a complex to perform H3K9 demethylation. The genome-wide regulation analysis revealed that OCT4 (O) and SOX2 (S), the core pluripotency transcriptional activators, maintained the pluripotent state of piPSCs depending on the H3K9 hypomethylation. Further investigation revealed that O/S cooperating with histone demethylase complex containing KDM3A and KDM3B promoted pluripotency 2 of 18 | ZHU et al.

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Generation of embryonic stem‐like cells from in vivo‐derived porcine blastocysts at a low concentration of basic fibroblast growth factor

Cited by 11 publications

References 40 publications

BCL2 enhances survival of porcine pluripotent stem cells through promoting FGFR2

BCL2 enhances survival of porcine pluripotent stem cells through promoting FGFR2

Effect of Optimized Concentrations of Basic Fibroblast Growth Factor and Epidermal Growth Factor on Proliferation of Fibroblasts and Expression of Collagen

Histone demethylase complexes KDM3A and KDM3B cooperate with OCT4/SOX2 to define a pluripotency gene regulatory network

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