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

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 Methods for Automated Single Cell Isolation and Sub‐Cloning of Human Pluripotent Stem Cells

2020 ◽  
Vol 55 (1) ◽  
Author(s):  
Valeria Fernandez Vallone ◽  
Narasimha Swamy Telugu ◽  
Iris Fischer ◽  
Duncan Miller ◽  
Sandra Schommer ◽  
...  
Keyword(s):  
Stem Cells ◽  
Single Cell ◽  
Pluripotent Stem Cells ◽  
Cell Isolation ◽  
Human Pluripotent Stem Cells ◽  
Single Cell Isolation

scholarly journals Revealing Cellular Heterogeneity and In Vitro Differentiation Trajectory of Cultured Human Endometrial Mesenchymal-like Stem Cells Using Single-cell RNA Sequencing

2020 ◽  
Author(s):  
Dandan Cao ◽  
Rachel W.S. Chan ◽  
Ernest H.Y. Ng ◽  
Kristina Gemzell-Danielsson ◽  
William S.B. Yeung
Keyword(s):  
Stem Cells ◽  
Single Cell ◽  
Rna Sequencing ◽  
Adult Stem Cells ◽  
Cellular Heterogeneity ◽  
In Vitro Differentiation ◽  
Regulatory Pathways ◽  
Single Cell Rna Sequencing ◽  
Unbiased Manner

ABSTRACTEndometrial mesenchymal-like stem cells (eMSCs) are adult stem cells contributing to endometrial regeneration. One set of perivascular markers (CD140b+CD146+) have been widely used to enrich eMSCs. Although eMSCs are easily accessible for regenerative medicine and have long been studied, their cellular heterogeneity and molecular program controlling their expansion and differentiation in vitro remains largely unclear. In this study, we applied 10X genomics single-cell RNA sequencing to eMSCs cultured in vitro after microbeading from 7 donors to investigate cellular heterogeneity in an unbiased manner. Corresponding clonogenic progenies of eMSCs after culture for 14 days were also sequenced to construct the in vitro differentiation trajectory of eMSCs. Transcriptomic expression based clustering revealed several subpopulations in eMSCs. Each subpopulation manifested distinct functional characteristics associated with immunomodulation, proliferation, extracellular matrix organization and cell differentiation. Pseudotime trajectory analysis on eMSCs and their differentiated progenies identified in vitro differentiation hierarchy of eMSCs. Further ligand-receptor pair analysis found that WNT signaling, NOTCH signaling, TGF-beta signaling and FGF signaling were important regulatory pathways for eMSC self-renewal and differentiation. By comparing eMSCs to Wharton’s Jelly MSCs and adipose-derived MSCs, we found these 3 kinds of MSCs expressed largely overlapping differentiation (CD) genes and highly variable genes. In summary, we reveal for the first time high molecular and cellular heterogeneity in cultured eMSCs, and identify the key signaling pathways that may be important for eMSC differentiation.

Epithelial In vitro Differentiation of Mesenchymal Stem Cells

2018 ◽  
Vol 13 (6) ◽  
pp. 409-422 ◽  
Author(s):  
Alvaro Sierra-Sanchez ◽  
Alexandra Ordonez-Luque ◽  
Olga Espinosa Ibanez ◽  
Antonio Ruiz-Garcia ◽  
Salvador Arias Santiago
Keyword(s):  
Stem Cells ◽  
Mesenchymal Stem Cells ◽  
In Vitro Differentiation

Mini Review; Differentiation of Human Pluripotent Stem Cells into Oocytes

2020 ◽  
Vol 15 (4) ◽  
pp. 301-307 ◽  
Author(s):  
Gaifang Wang ◽  
Maryam Farzaneh
Keyword(s):  
Stem Cells ◽  
Pluripotent Stem Cells ◽  
Embryonic Stem ◽  
Human Pluripotent Stem Cells ◽  
Human Oocyte ◽  
Differentiation Potential ◽  
Cell Lineages ◽  
Cell Based Therapy ◽  
Induced Pluripotent

Primary Ovarian Insufficiency (POI) is one of the main diseases causing female infertility that occurs in about 1% of women between 30-40 years of age. There are few effective methods for the treatment of women with POI. In the past few years, stem cell-based therapy as one of the most highly investigated new therapies has emerged as a promising strategy for the treatment of POI. Human pluripotent stem cells (hPSCs) can self-renew indefinitely and differentiate into any type of cell. Human Embryonic Stem Cells (hESCs) as a type of pluripotent stem cells are the most powerful candidate for the treatment of POI. Human-induced Pluripotent Stem Cells (hiPSCs) are derived from adult somatic cells by the treatment with exogenous defined factors to create an embryonic-like pluripotent state. Both hiPSCs and hESCs can proliferate and give rise to ectodermal, mesodermal, endodermal, and germ cell lineages. After ovarian stimulation, the number of available oocytes is limited and the yield of total oocytes with high quality is low. Therefore, a robust and reproducible in-vitro culture system that supports the differentiation of human oocytes from PSCs is necessary. Very few studies have focused on the derivation of oocyte-like cells from hiPSCs and the details of hPSCs differentiation into oocytes have not been fully investigated. Therefore, in this review, we focus on the differentiation potential of hPSCs into human oocyte-like cells.

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