scholarly journals Modeling Hypoxic Stress In Vitro Using Human Embryonic Stem Cells Derived Cardiomyocytes Matured by FGF4 and Ascorbic Acid Treatment

Cells ◽  
2021 ◽  
Vol 10 (10) ◽  
pp. 2741
Author(s):  
Seung-Cheol Choi ◽  
Ha-Rim Seo ◽  
Long-Hui Cui ◽  
Myeong-Hwa Song ◽  
Ji-Min Noh ◽  
...  
Keyword(s):  
Stem Cells ◽  
Ascorbic Acid ◽  
Human Embryonic Stem Cell ◽  
Culture Medium ◽  
Embryonic Stem ◽  
Cardiac Biomarkers ◽  
Hypoxic Stress ◽  
Hypoxic Injury ◽  
Diagnosis And Prognosis

Mature cardiomyocytes (CMs) obtained from human pluripotent stem cells (hPSCs) have been required for more accurate in vitro modeling of adult-onset cardiac disease and drug discovery. Here, we found that FGF4 and ascorbic acid (AA) induce differentiation of BG01 human embryonic stem cell–cardiogenic mesoderm cells (hESC-CMCs) into mature and ventricular CMs. Co-treatment of BG01 hESC-CMCs with FGF4+AA synergistically induced differentiation into mature and ventricular CMs. FGF4+AA-treated BG01 hESC-CMs robustly released acute myocardial infarction (AMI) biomarkers (cTnI, CK-MB, and myoglobin) into culture medium in response to hypoxic injury. Hypoxia-responsive genes and potential cardiac biomarkers proved in the diagnosis and prognosis of coronary artery diseases were induced in FGF4+AA-treated BG01 hESC-CMs in response to hypoxia based on transcriptome analyses. This study demonstrates that it is feasible to model hypoxic stress in vitro using hESC-CMs matured by soluble factors.

scholarly journals Blockade of STAT3 Causes Severe In Vitro and In Vivo Maturation Defects in Intestinal Organoids Derived from Human Embryonic Stem Cells

2019 ◽  
Vol 8 (7) ◽  
pp. 976 ◽  
Author(s):  
Kwang Bo Jung ◽  
Ohman Kwon ◽  
Mi-Ok Lee ◽  
Hana Lee ◽  
Ye Seul Son ◽  
...  
Keyword(s):  
Stem Cells ◽  
Human Embryonic Stem Cell ◽  
In Vitro Maturation ◽  
Embryonic Stem ◽  
Generating Functional ◽  
Intestinal Maturation ◽  
Intestinal Organoids ◽  
Hesc Lines

Human intestinal organoids (hIOs), which resemble the human intestine structurally and physiologically, have emerged as a new modality for the study of the molecular and cellular biology of the intestine in vitro. We recently developed an in vitro maturation technique for generating functional hIOs from human pluripotent stem cells (hPSCs). Here, we investigated the function of STAT3 for inducing in vitro maturation of hIOs. This was accompanied by the tyrosine phosphorylation of STAT3, whereas treatment with pharmacological inhibitors of STAT3 suppressed the phosphorylation of STAT3 and the expression of intestinal maturation markers. We generated and characterized STAT3 knockout (KO) human embryonic stem cell (hESC) lines using CRISPR/Cas9-mediated gene editing. We found that STAT3 KO does not affect the differentiation of hESCs into hIOs but rather affects the in vitro maturation of hIOs. STAT3 KO hIOs displayed immature morphologies with decreased size and reduced budding in hIOs even after in vitro maturation. STAT3 KO hIOs showed markedly different profiles from hIOs matured in vitro and human small intestine. Additionally, STAT3 KO hIOs failed to maintain upon in vivo transplantation. This study reveals a core signaling pathway consisting of STAT3 controlling the in vitro maturation of hIOs derived from hPSCs.

scholarly journals Biofunctionalised bacterial cellulose scaffold supports the patterning and expansion of human embryonic stem cell-derived dopaminergic progenitor cells

2021 ◽  
Vol 12 (1) ◽  
Author(s):  
Miranda Robbins ◽  
Venkat Pisupati ◽  
Roberta Azzarelli ◽  
Samer I. Nehme ◽  
Roger A. Barker ◽  
...  
Keyword(s):  
Parkinson’S Disease ◽  
Stem Cells ◽  
Parkinson's Disease ◽  
Stem Cell ◽  
Embryonic Stem Cell ◽  
Bacterial Cellulose ◽  
Progenitor Cells ◽  
Human Embryonic Stem Cell ◽  
Embryonic Stem

Abstract Background Stem cell-based therapies for neurodegenerative diseases like Parkinson’s disease are a promising approach in regenerative medicine and are now moving towards early stage clinical trials. However, a number of challenges remain including the ability to grow stem cells in vitro on a 3-dimensional scaffold, as well as their loss, by leakage or cell death, post-implantation. These issues could, however, be helped through the use of scaffolds that support the growth and differentiation of stem cells both in vitro and in vivo. The present study focuses on the use of bacterial cellulose as an in vitro scaffold to promote the growth of different stem cell-derived cell types. Bacterial cellulose was used because of its remarkable properties such as its wettability, ability to retain water and low stiffness, all of which is similar to that found in brain tissue. Methods We cultured human embryonic stem cell-derived progenitor cells on bacterial cellulose with growth factors that were covalently functionalised to the surface via silanisation. Epifluorescence microscopy and immunofluorescence were used to detect the differentiation of stem cells into dopaminergic ventral midbrain progenitor cells. We then quantified the proportion of cells that differentiated into progenitor cells and compared the effect of growing cells on biofunctionalised cellulose versus standard cellulose. Results We show that the covalent functionalisation of bacterial cellulose sheets with bioactive peptides improves the growth and differentiation of human pluripotent stem cells into dopaminergic neuronal progenitors. Conclusions This study suggests that the biocompatible material, bacterial cellulose, has potential applications in cell therapy approaches as a means to repair damage to the central nervous system, such as in Parkinson’s disease but also in tissue engineering.

scholarly journals Stem Cells, Sex, and Procreation

2003 ◽  
Vol 12 (4) ◽  
pp. 353-371 ◽  
Author(s):  
JOHN HARRIS
Keyword(s):  
Stem Cells ◽  
Human Embryonic Stem Cell ◽  
Embryonic Stem ◽  
Fetal Tissue ◽  
Ethical Principle ◽  
Embryo Research ◽  
Vitro Fertilization ◽  
Heterosexual Intercourse ◽  
Human Embryo Research

Sex is not the answer to everything, though young men think it is, but it may be the answer to the intractable debate over the ethics of human embryonic stem cell research. In this paper, I advance one ethical principle that, as yet, has not received the attention its platitudinous character would seem to merit. If found acceptable, this principle would permit the beneficial use of any embryonic or fetal tissue that would, by default, be lost or destroyed. More important, I make two appeals to consistency, or to parity of reasoning, that I believe show that no one who either has used or intends to use sexual reproduction as their means of procreation, nor indeed anyone who has unprotected heterosexual intercourse, nor anyone who finds in vitro fertilization (IVF) acceptable, nor anyone who believes that abortion is ever permissible can consistently object on principle to human embryo research nor to the use of embryonic stem cells for research or therapy.

scholarly journals Role of TGFβ1 and WNT6 in FGF2 and BMP4-driven endothelial differentiation of murine embryonic stem cells

Angiogenesis ◽  
2021 ◽  
Author(s):  
Anna Gualandris ◽  
Alessio Noghero ◽  
Davide Cora’ ◽  
Elena Astanina ◽  
Marco Arese ◽  
...  
Keyword(s):  
Stem Cells ◽  
Embryonic Stem Cells ◽  
Culture Medium ◽  
Expression Profiles ◽  
Es Cells ◽  
Embryonic Stem ◽  
Endothelial Differentiation ◽  
Loss Of Function ◽  
Pure Cultures

AbstractEmbryonic stem cells (ES) are a valuable source of endothelial cells. By co-culturing ES cells with the stromal PA6 cells, the endothelial commitment can be achieved by adding exogenous FGF2 or BMP4. In this work, the molecular pathways that direct the differentiation of ES cells toward endothelium in response to FGF2 are evaluated and compared to those activated by BMP4. To this purpose the genes expression profiles of both ES/PA6 co-cultures and of pure cultures of PA6 cells were obtained by microarray technique at different time points. The bioinformatics processing of the data indicated TGFβ1 as the most represented upstream regulator in FGF2-induced endothelial commitment while WNT pathway as the most represented in BMP4-activated endothelial differentiation. Loss of function experiments were performed to validate the importance of TGFβ1 and WNT6 respectively in FGF2 and BMP4-induced endothelial differentiation. The loss of TGFβ1 expression significantly impaired the accomplishment of the endothelial commitment unless exogenous recombinant TGFβ1 was added to the culture medium. Similarly, silencing WNT6 expression partially affected the endothelial differentiation of the ES cells upon BMP4 stimulation. Such dysfunction was recovered by the addition of recombinant WNT6 to the culture medium. The ES/PA6 co-culture system recreates an in vitro complete microenvironment in which endothelial commitment is accomplished in response to alternative signals through different mechanisms. Given the importance of WNT and TGFβ1 in mediating the crosstalk between tumor and stromal cells this work adds new insights in the mechanism of tumor angiogenesis and of its possible inhibition.

scholarly journals Similar Population of CD133+ and DDX4+ VSEL-Like Stem Cells Sorted from Human Embryonic Stem Cell, Ovarian, and Ovarian Cancer Ascites Cell Cultures: The Real Embryonic Stem Cells?

Cells ◽  
2019 ◽  
Vol 8 (7) ◽  
pp. 706 ◽  
Author(s):  
Irma Virant-Klun ◽  
Petra Skerl ◽  
Srdjan Novakovic ◽  
Eda Vrtacnik-Bokal ◽  
Spela Smrkolj
Keyword(s):  
Ovarian Cancer ◽  
Stem Cells ◽  
Stem Cell ◽  
Embryonic Stem Cells ◽  
Embryonic Stem Cell ◽  
Cell Cultures ◽  
Human Embryonic Stem Cell ◽  
Embryonic Stem ◽  
Recurrent Ovarian Cancer

A population of small stem cells with diameters of up to 5 μm resembling very small embryonic-like stem cells (VSELs) were sorted from human embryonic stem cell (hESC) cultures using magnetic-activated cell sorting (MACS) based on the expression of a stem-cell-related marker prominin-1 (CD133). These VSEL-like stem cells had nuclei that almost filled the whole cell volume and expressed stem-cell-related markers (CD133, SSEA-4) and markers of germinal lineage (DDX4/VASA, PRDM14). They were comparable to similar populations of small stem cells sorted from cell cultures of normal ovaries and were the predominant cells in ascites of recurrent ovarian cancer. The sorted populations of CD133+ VSEL-like stem cells were quiescent in vitro, except for ascites, and were highly activated after exposure to valproic acid and follicle-stimulating hormone (FSH), indicating a new tool to study these cells in vitro. These VSEL-like stem cells spontaneously formed clusters resembling tumour-like structures or grew into larger, oocyte-like cells and were differentiated in vitro into adipogenic, osteogenic and neural lineages after sorting. We propose the population of VSEL-like stem cells from hESC cultures as potential original embryonic stem cells, which are present in the human embryo, persist in adult human ovaries from the embryonic period of life and are involved in cancer manifestation.

scholarly journals Characterization of human embryonic stem cells in animal component-free medium

2020 ◽  
Author(s):  
Masakazu Machida ◽  
Rie Abutani ◽  
Hiroshi Miyajima ◽  
Tetsuji Sasaki ◽  
Yoshiko Abe ◽  
...  
Keyword(s):  
Stem Cells ◽  
Embryonic Stem Cells ◽  
Human Embryonic Stem Cells ◽  
Culture Medium ◽  
Embryonic Stem ◽  
Good Manufacturing Practice ◽  
Raw Material ◽  
Population Doublings ◽  
Animal Component

Clinical use of human embryonic stem cells (ESCs) as a raw material requires good manufacturing practice-compliant axillary materials such as culture medium. To this end, animal components should not be used and contamination of virus/bacteria/fungus and allergens are a concern. In addition, animal components such as albumin and fetal bovine serum pose difficulties such as a lot-to-lot variation. However, only a limited number of animal component-free media have been developed to date. In this study, we investigated whether SEES2 ESCs can be stably propagated for 16 passages (54 population doublings) over a period of 60 days in a newly established Stem-Partner ACF medium. SEES2 ESC maintained their intact karyotype, i.e. 46,XX, and their undifferentiated phenotypes after long-term culture. An in vitro differentiation assay revealed that SEES2 ESCs exhibited multipotency, i.e. endodermal, mesodermal and ectodermal differentiation. Subcutaneous implantation of SEES2 ESCs generated mature teratomas without malignant transformation. These results show that SEES2 ESCs in the Stem-Partner ACF medium can be used to establish master cell banks for future regenerative medicine as well as other ESCs in the previously reported culture medium.

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