A screen of suitable inducers for germline differentiation of chicken embryonic stem cells

Shi, Qingqing; Sun, Min; Zhang, Zhentao; Zhang, Yani; Elsayed, Ahmed K.; Zhang, Lei; Huang, Xiaomei; Li, Bichun

doi:10.1016/j.anireprosci.2014.04.002

Cited by 19 publications

(23 citation statements)

References 45 publications

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“…The extracellular matrix and basement membrane constitute the main niche for germ cell as well as sertoli and interstitial cell that support their functions [ 9 ]. A culture system contained laminin and fibronectin, but not collagen along with meiosis inducer, and sertoli cell co-culture led to an elevation in the spermatogonia stem cell marker expression such as DAZL by chick embryonic stem cells [ 10 ]. Glycosaminoglycans (GAG) such as heparin sulfate facilitated the primordial germ cell migration and survival [ 11 ].…”

Section: Introductionmentioning

confidence: 99%

Characterization, recellularization, and transplantation of rat decellularized testis scaffold with bone marrow-derived mesenchymal stem cells

et al. 2018

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BackgroundRegenerative medicine potentially offers the opportunity for curing male infertility. Native extracellular matrix (ECM) creates a reconstruction platform to replace the organs. In this study, we aimed to evaluate the efficiency of the testis decellularized scaffold as a proper niche for stem cell differentiation toward testis-specific cell lineages.MethodsRats’ testes were decellularized by freeze-thaw cycle followed by immersion in deionized distilled water for 2 h, perfused with 1% Triton X-100 through ductus deferens for 4 h, 1% SDS for 48 h and 1% DNase for 2 h. The decellularized samples were prepared for further in vitro and in vivo analyses.ResultHistochemical and immunohistochemistry studies revealed that ECM components such as Glycosaminoglycans (GAGs), neutral carbohydrate, elastic fibers, collagen I & IV, laminin, and fibronectin were well preserved, and the cells were completely removed after decellularization. Scanning electron microscopy (SEM) showed that 3D ultrastructure of the testis remained intact. In vivo and in vitro studies point out that decellularized scaffold was non-toxic and performed a good platform for cell division. In vivo implant of the scaffolds with or without mesenchymal stem cells (MSCs) showed that appropriate positions for transplantation were the mesentery and liver and the scaffolds could induce donor-loaded MSCs or host migrating cells to differentiate to the cells with phenotype of the sertoli- and leydig-like cells. The scaffolds also provide a good niche for migrating DAZL-positive cells; however, they could not differentiate into post meiotic-cell lineages.ConclusionThe decellularized testis can be considered as a promising vehicle to support cell transplantation and may provide an appropriate niche for testicular cell differentiation.

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Section: Introductionmentioning

confidence: 99%

Characterization, recellularization, and transplantation of rat decellularized testis scaffold with bone marrow-derived mesenchymal stem cells

et al. 2018

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“…To study the function of C1EIP in PGC generation, we implemented a model for inducing ESC differentiation into SSCs by in vitro RA treatment described previously by our lab ( Figure 3A ) ( Shi et al, 2014 ). C1EIP was knocked down (shRNA) and overexpressed using lentiviruses vector ( Figure 3B and Supplementary Figure S3 ).…”

Section: Resultsmentioning

confidence: 99%

“…The PGC induction of the retinoic acid (RA) model was established following a previously described method ( Shi et al, 2014 ). In brief, the chicken ESCs (5 × 10 4 cell/well) were cultured in 24-well plates in DMEM medium supplemented with 10 mM RA (Solarbio, Beijing, China, IR0060), 15% FBS (Gibco, New York, NY, United States, 26140), and fibronectin (5 μg/cm 2 ).…”

Section: Methodsmentioning

confidence: 99%

C1EIP Functions as an Activator of ENO1 to Promote Chicken PGCs Formation via Inhibition of the Notch Signaling Pathway

et al. 2020

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The production of germ cells, especially primordial germ cells (PGCs), is important for avian stem cells and reproduction biology. However, key factors involved in the regulation of PGCs remain unknown. Here, we report a PGC-related marker gene: C1EIP (Chromosome 1 Expression in PGCs), whose activation and expression are regulated by the transcription factor STAT3 (signal transducer and activator of transcription 3), histone acetylation, and promoter methylation. C1EIP regulates PGCs formation by mediating the expression of PGC-associated genes, such as CVH (Chicken Vasa Homologous) and CKIT (Chicken KIT proto-oncogene). C1EIP knockdown during embryonic development reduces PGC generation efficiency both in vitro and in ovo . Conversely, C1EIP overexpression increases the formation efficiency of PGCs. C1EIP encodes a cytoplasmic protein that interacts with ENO1 (Enolase 1) in the cytoplasm, inhibits the Notch signaling pathway, and positively regulates PGC generation. Collectively, our findings demonstrate C1EIP as a novel gene involved in PGC formation, which regulates genes involved in embryonic stem cell differentiation through interaction with ENO1 and subsequent inhibition of the Notch signaling pathway by the impression of Myc (MYC proto-oncogene).

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“…Dazl , an intrinsic meiotic competence factor, is required for Stra8 -mediated initiation of meiosis in germ cells ( Lin et al, 2008 ). Overexpression of Stra8 and Dazl genes promotes the transdifferentiation of mesenchymal stem cells and ESCs in vitro toward PGCs ( Li et al, 2017 ; Shi et al, 2014 ). The elevated expression of Pou5f1 , Dazl , Stella and Stra8 might indicate that the GFP positive cells separated from teratomas partially develop towards germ cells.…”

Section: Discussionmentioning

confidence: 99%

Murine pluripotent stem cells that escape differentiation inside teratomas maintain pluripotency

Pei

Yue

Zhang

et al. 2018

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BackgroundPluripotent stem cells (PSCs) offer immense potential as a source for regenerative therapies. The teratoma assay is widely used in the field of stem cells and regenerative medicine, but the cell composition of teratoma is still elusive.MethodsWe utilized PSCs expressing enhanced green fluorescent protein (EGFP) under the control of the Pou5f1 promoter to study the persistence of potential pluripotent cells during teratoma formation in vivo. OCT4-MES (mouse embryonic stem cells) were isolated from the blastocysts of 3.5-day OCT4-EGFP mice (transgenic mice express EGFP cDNA under the control of the Pou5f1 promoter) embryos, and TG iPS 1-7 (induced pluripotent stem cells) were generated from mouse embryonic fibroblasts (MEFs) from 13.5-day OCT4-EGFP mice embryos by infecting them with a virus carrying OCT4, SOX2, KLF4 and c-MYC. These pluripotent cells were characterized according to their morphology and expression of pluripotency markers. Their differentiation ability was studied with in vivo teratoma formation assays. Further differences between pluripotent cells were examined by real-time quantitative PCR (qPCR).ResultsThe results showed that several OCT4-expressing PSCs escaped differentiation inside of teratomas, and these escaped cells (MES-FT, GFP-positive cells separated from OCT4-MES-derived teratomas; and iPS-FT, GFP-positive cells obtained from teratomas formed by TG iPS 1-7) retained their pluripotency. Interestingly, a small number of GFP-positive cells in teratomas formed by MES-FT and iPS-FT (MES-ST, GFP-positive cells isolated from MES-FT-derived teratomas; iPS-ST, GFP-positive cells obtained from teratomas formed by iPS-FT) were still pluripotent, as shown by alkaline phosphatase (AP) staining, immunofluorescent staining and PCR. MES-FT, iPS-FT, MES-ST and iPS-ST cells also expressed several markers associated with germ cell formation, such as Dazl, Stella and Stra8.ConclusionsIn summary, a small number of PSCs escaped differentiation inside of teratomas, and these cells maintained pluripotency and partially developed towards germ cells. Both escaped PSCs and germ cells present a risk of tumor formation. Therefore, medical workers must be careful in preventing tumor formation when stem cells are used to treat specific diseases.

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A screen of suitable inducers for germline differentiation of chicken embryonic stem cells

Cited by 19 publications

References 45 publications

Characterization, recellularization, and transplantation of rat decellularized testis scaffold with bone marrow-derived mesenchymal stem cells

Characterization, recellularization, and transplantation of rat decellularized testis scaffold with bone marrow-derived mesenchymal stem cells

C1EIP Functions as an Activator of ENO1 to Promote Chicken PGCs Formation via Inhibition of the Notch Signaling Pathway

Murine pluripotent stem cells that escape differentiation inside teratomas maintain pluripotency

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