scholarly journals Studying human reproductive biology through single-cell analysis and in vitro differentiation of stem cells into germ cell-like cells

2020 ◽  
Vol 26 (5) ◽  
pp. 670-688 ◽  
Author(s):  
Lin Li ◽  
Risako Yang ◽  
Chenghong Yin ◽  
Kehkooi Kee
Keyword(s):  
Stem Cells ◽  
Single Cell ◽  
Germ Cells ◽  
Regulatory Networks ◽  
Single Cell Analysis ◽  
Reproductive Development ◽  
In Vitro Differentiation ◽  
Gene Markers ◽  
Human Reproductive

Abstract BACKGROUND Understanding the molecular and cellular mechanisms of human reproductive development has been limited by the scarcity of human samples and ethical constraints. Recently, in vitro differentiation of human pluripotent stem cells into germ cells and single-cell analyses have opened new avenues to directly study human germ cells and identify unique mechanisms in human reproductive development. OBJECTIVE AND RATIONALE The goal of this review is to collate novel findings and insightful discoveries with these new methodologies, aiming at introducing researchers and clinicians to the use of these tools to study human reproductive biology and develop treatments for infertility. SEARCH METHODS PubMed was used to search articles and reviews with the following main keywords: in vitro differentiation, human stem cells, single-cell analysis, spermatogenesis, oogenesis, germ cells and other key terms related to these subjects. The search period included all publications from 2000 until now. OUTCOMES Single-cell analyses of human gonads have identified many important gene markers at different developmental stages and in subpopulations of cells. To validate the functional roles of these gene markers, researchers have used the in vitro differentiation of human pluripotent cells into germ cells and confirmed that some genetic requirements are unique in human germ cells and are not conserved in mouse models. Moreover, transcriptional regulatory networks and the interaction of germ and somatic cells in gonads were elucidated in these studies. WIDER IMPLICATIONS Single-cell analyses allow researchers to identify gene markers and potential regulatory networks using limited clinical samples. On the other hand, in vitro differentiation methods provide clinical researchers with tools to examine these newly identify gene markers and study the causative effects of mutations previously associated with infertility. Combining these two methodologies, researchers can identify gene markers and networks which are essential and unique in human reproductive development, thereby producing more accurate diagnostic tools for assessing reproductive disorders and developing treatments for infertility.

scholarly journals scATAC-Seq reveals epigenetic heterogeneity associated with an EMT-like process in male germline stem cells and its regulation by G9a

2020 ◽  
Author(s):  
Jinyue Liao ◽  
Hoi Ching Suen ◽  
Alfred Chun Shui Luk ◽  
Annie Wing Tung Lee ◽  
Judy Kin Wing Ng ◽  
...  
Keyword(s):  
Stem Cells ◽  
Single Cell ◽  
Regulatory Networks ◽  
Epithelial Mesenchymal Transition ◽  
Stem Cell Differentiation ◽  
Germline Stem Cells ◽  
Gene Markers ◽  
Mesenchymal Transition

AbstractBackgroundEpithelial-mesenchymal transition (EMT) is a phenomenon in which epithelial cells acquire mesenchymal traits. It contributes to organogenesis and tissue homeostasis, as well as stem cell differentiation. Emerging evidence indicates that heterogeneous expression of EMT gene markers presents in sub-populations of germline stem cells (GSCs). However, the functional implications of such heterogeneity are largely elusive.ResultsWe unravelled an EMT-like process in GSCs by in vitro extracellular matrix (ECM) model and single-cell genomics approaches. We found that histone methyltransferase G9a regulated an EMT-like program in GSC in vitro and contributed to neonatal germ cell migration in vivo. Through modulating ECM, we demonstrated that GSCs exist in interconvertible epithelial-like and mesenchymal-like cell states. GSCs gained higher migratory ability after transition to a mesenchymal-like cell state, which was largely mediated by the TGF-β signaling pathway. Dynamics of epigenetic regulation at the single-cell level was also found to align with the EMT-like process. Chromatin accessibility profiles generated by single-cell sequencing assay for transposase-accessible chromatin (scATAC-seq) clustered GSCs into epithelial-like and mesenchymal-like states, which were associated with differentiation status. The high-resolution data revealed regulators in the EMT-like process, including transcription factors Zeb1. We further identified putative enhancer-promoter interactions and cis-co-accessibility networks at loci such as Tgfb1, Notch1 and Lin28a. Lastly, we identified HES1 as the putative target underlying G9a’s regulation.ConclusionOur work provides the foundation for understanding the EMT-like process and a comprehensive resource for future investigation of epigenetic regulatory networks in GSCs.

In vitro differentiation of germ cells from stem cells

2013 ◽  
pp. 236-249
Author(s):  
Fumihiro Sugawa ◽  
Karin Hübner ◽  
Hans R. Schöler
Keyword(s):  
Stem Cells ◽  
Germ Cells ◽  
In Vitro Differentiation

scholarly journals In vitro differentiation of human oocyte-like cells from oogonial stem cells: single-cell isolation and molecular characterization

2018 ◽  
Vol 33 (3) ◽  
pp. 464-473 ◽  
Author(s):  
Erica Silvestris ◽  
Paola Cafforio ◽  
Stella D’Oronzo ◽  
Claudia Felici ◽  
Franco Silvestris ◽  
...  
Keyword(s):  
Stem Cells ◽  
Single Cell ◽  
Molecular Characterization ◽  
Cell Isolation ◽  
Human Oocyte ◽  
In Vitro Differentiation ◽  
Single Cell Isolation

In vitro Differentiation of Germ Cells from Stem Cells

2016 ◽  
Vol 9 (4) ◽  
pp. 305-310 ◽  
Author(s):  
Xinbao Ding ◽  
Jian Wang ◽  
Ji Wu
Keyword(s):  
Stem Cells ◽  
Germ Cells ◽  
In Vitro Differentiation

Establishment of mouse male germ line stem cells (GSCs) and in vitro differentiation to haploid germ cells

2003 ◽  
Vol 80 ◽  
pp. 1
Author(s):  
D.R. Lee ◽  
S.K. Kim ◽  
K.Y. Cha ◽  
Y.H. Yang ◽  
T.K. Yoon ◽  
...  
Keyword(s):  
Stem Cells ◽  
Germ Cells ◽  
Germ Line ◽  
In Vitro Differentiation ◽  
Male Germ Line

Abstract P353: Adipose-derived Stem Cells Contribute To Vascular Remodeling Via Clec11a + Subpopulation

2021 ◽  
Vol 129 (Suppl_1) ◽  
Author(s):  
Yongli Ji ◽  
Yunrui Lu ◽  
Jian Shen ◽  
Meixiang Xiang ◽  
Yao Xie
Keyword(s):  
Stem Cells ◽  
Single Cell ◽  
Vascular Remodeling ◽  
Single Cell Analysis ◽  
Gene Set Enrichment Analysis ◽  
Adipose Derived Stem Cells ◽  
Cell Analysis ◽  
Tgf Β1

Introduction: Recent researches identified the existence of perivascular adipose-derived stem cells (ADSCs) which could differentiate into vascular lineages and participate in vascular remodeling. Single-cell mRNA analysis revealed cellular heterogeneity of subcutaneous ADSCs in respect to cell clustering and cell differentiation. However, such analysis of perivascular ADSCs has not been investigated at a single-cell level. Hypothesis: There is a significant difference among perivascular ADSCs subpopulations in respect to vascular-lineage differentiation. Methods: We performed droplet-based single-cell profiling of subcutaneous and perivascular adipose stromal cells and compared ADSCs regarding their heterogeneity, gene ontology, and cell fate trajectory by applying single-cell analysis as well as in vitro and in vivo assays. Results: Single-cell analysis uncovered 4 perivascular ADSCs subpopulations including Dpp4+ , Col4a2+ / Icam1+ , Clec11a+ / Cpe+ and Sult1e1+ cells. Notably, the Clec11a + subpopulation comprised the bulk of perivascular ADSCs, while was hardly presented in subcutaneous ADSCs. Further gene-set enrichment analysis suggested Clec11a + ADSCs were potentially involved with TGF-β signaling pathways and pseudotemporal analysis predicted that Clec11a + subpopulation lay at the end of the differential trajectory towards smooth muscle cells (SMCs). In vitro assays displayed that perivascular ADSCs could differentiate into SMCs via CLEC11A regulation when treated by TGF-β1. To further elucidate the role of the Clec11a + subpopulation in SMCs differentiation, we labeled CLEC11A+ and CLEC11A- perivascular ADSCs by lentivirus transfection and isolated them by FACS assay. CLEC11A+ cells showed the greater capability of SMCs differentiation in response to TGF-β1 in vitro and enhanced neointima formation when transplanted to the adventitial side of guidewire injured arteries. Conclusions: The present study depicted the unique heterogeneity of perivascular ADSCs and the novel role of the Clec11a + subpopulation, providing a supplement for the relationship between perivascular ADSCs and vascular SMCs.

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