Oxygen as a Master Regulator of Human Pluripotent Stem Cell Function and Metabolism

Human-induced pluripotent stem cells (hiPSCs) offer numerous possibilities in science and medicine, particularly when combined with precise genome editing methods. hiPSCs are artificially generated equivalents of human embryonic stem cells (hESCs), which possess an unlimited ability to self-renew and the potential to differentiate into any cell type of the human body. Importantly, generating patient-specific hiPSCs enables personalized drug testing or autologous cell therapy upon differentiation into a desired cell line. However, to ensure the highest standard of hiPSC-based biomedical products, their safety and reliability need to be proved. One of the key factors influencing human pluripotent stem cell (hPSC) characteristics and function is oxygen concentration in their microenvironment. In recent years, emerging data have pointed toward the beneficial effect of low oxygen pressure (hypoxia) on both hiPSCs and hESCs. In this review, we examine the state-of-the-art research on the oxygen impact on hiPSC functions and activity with an emphasis on their niche, metabolic state, reprogramming efficiency, and differentiation potential. We also discuss the similarities and differences between PSCs and cancer stem cells (CSCs) with respect to the role of oxygen in both cell types.

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Abstract 11: Xenogen-free In Vitro Differentiation and Expansion of Human Pluripotent Stem Cell-Derived Cardiovascular Progenitor Cells

Circulation Research ◽

10.1161/res.115.suppl_1.11 ◽

2014 ◽

Vol 115 (suppl_1) ◽

Author(s):

Xianmei Meng ◽

Anne Knopf ◽

Agustin Vega-Crespo ◽

James Byrne ◽

Ben Van Handel ◽

...

Keyword(s):

Stem Cells ◽

Stem Cell ◽

Progenitor Cells ◽

Pluripotent Stem Cell ◽

Single Cells ◽

Embryonic Stem ◽

Human Pluripotent Stem Cell ◽

Differentiation Potential ◽

Cell Based Therapy ◽

Trilineage Differentiation

Background: Human pluripotent stem cell-derived cardiovascular progenitor cells (hPSC-CPCs) represent a tractable option for cell-based therapy for heart disease. However, to be clinically relevant, these cells must be derived under good manufacturing practices (GMP)-compatible conditions and produced in great enough quantities to treat adult patients. Here we sought to demonstrate for the first time the generation and expansion of clinically relevant numbers of hPSC-CPCs in xenogen-free protocol. Methods and Results: GMP-grade human induced pluripotent stem cells (GMP-hiPSCs) and human embryonic stem cells (H1 and H9) were dissociated into single cells and cultured in low attachment dishes to differentiate into CPCs in StemPro medium including small molecules and human cytokines with high efficiency of 86%, 80% and 66% for GMP-hiPSCs, H1 and H9, respectively (Figure 1). All hPSC-CPCs possessed trilineage differentiation potentials, as shown by differentiation into endothelial and smooth muscle cells and functional cardiomyocytes (Figure 2). Moreover, sorted hPSC-CPCs expanded >5 fold in 10 days in xenogen-free conditions while still maintaining trilineage differentiation potential and an efficiency of ~70% (Figure 3). Conclusions: Here we demonstrate a xenogeny-free CPC derivation and expansion protocol that can generate clinically relevant numbers of GMP-grade cardiovascular progenitors that could be used in a clinical setting.

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Multiple Roles for Cholinergic Signaling from the Perspective of Stem Cell Function

International Journal of Molecular Sciences ◽

10.3390/ijms22020666 ◽

2021 ◽

Vol 22 (2) ◽

pp. 666

Author(s):

Toshio Takahashi

Keyword(s):

Stem Cells ◽

Stem Cell ◽

Cell Function ◽

Cell Activity ◽

Embryonic Stem ◽

Cholinergic Neurons ◽

Cell Types ◽

Proliferative Potential ◽

True Nature ◽

Cholinergic Signaling

Stem cells have extensive proliferative potential and the ability to differentiate into one or more mature cell types. The mechanisms by which stem cells accomplish self-renewal provide fundamental insight into the origin and design of multicellular organisms. These pathways allow the repair of damage and extend organismal life beyond that of component cells, and they probably preceded the evolution of complex metazoans. Understanding the true nature of stem cells can only come from discovering how they are regulated. The concept that stem cells are controlled by particular microenvironments, also known as niches, has been widely accepted. Technical advances now allow characterization of the zones that maintain and control stem cell activity in several organs, including the brain, skin, and gut. Cholinergic neurons release acetylcholine (ACh) that mediates chemical transmission via ACh receptors such as nicotinic and muscarinic receptors. Although the cholinergic system is composed of organized nerve cells, the system is also involved in mammalian non-neuronal cells, including stem cells, embryonic stem cells, epithelial cells, and endothelial cells. Thus, cholinergic signaling plays a pivotal role in controlling their behaviors. Studies regarding this signal are beginning to unify our understanding of stem cell regulation at the cellular and molecular levels, and they are expected to advance efforts to control stem cells therapeutically. The present article reviews recent findings about cholinergic signaling that is essential to control stem cell function in a cholinergic niche.

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Stem cell therapies and benefaction of somatic cell nuclear transfer cloning in COVID-19 era

Stem Cell Research & Therapy ◽

10.1186/s13287-021-02334-5 ◽

2021 ◽

Vol 12 (1) ◽

Author(s):

Birbal Singh ◽

Gorakh Mal ◽

Vinod Verma ◽

Ruchi Tiwari ◽

Muhammad Imran Khan ◽

...

Keyword(s):

Stem Cells ◽

Stem Cell ◽

Regenerative Medicine ◽

Somatic Cell ◽

Somatic Cell Nuclear Transfer ◽

Nuclear Transfer ◽

Drug Testing ◽

Human Interaction ◽

Patient Specific ◽

Wide Range

Abstract Background The global health emergency of COVID-19 has necessitated the development of multiple therapeutic modalities including vaccinations, antivirals, anti-inflammatory, and cytoimmunotherapies, etc. COVID-19 patients suffer from damage to various organs and vascular structures, so they present multiple health crises. Mesenchymal stem cells (MSCs) are of interest to treat acute respiratory distress syndrome (ARDS) caused by SARS-CoV-2 infection. Main body Stem cell-based therapies have been verified for prospective benefits in copious preclinical and clinical studies. MSCs confer potential benefits to develop various cell types and organoids for studying virus-human interaction, drug testing, regenerative medicine, and immunomodulatory effects in COVID-19 patients. Apart from paving the ways to augment stem cell research and therapies, somatic cell nuclear transfer (SCNT) holds unique ability for a wide range of health applications such as patient-specific or isogenic cells for regenerative medicine and breeding transgenic animals for biomedical applications. Being a potent cell genome-reprogramming tool, the SCNT has increased prominence of recombinant therapeutics and cellular medicine in the current era of COVID-19. As SCNT is used to generate patient-specific stem cells, it avoids dependence on embryos to obtain stem cells. Conclusions The nuclear transfer cloning, being an ideal tool to generate cloned embryos, and the embryonic stem cells will boost drug testing and cellular medicine in COVID-19.

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Cardiotoxicity evaluation using human embryonic stem cells and induced pluripotent stem cell-derived cardiomyocytes

Stem Cell Research & Therapy ◽

10.1186/s13287-017-0473-x ◽

2017 ◽

Vol 8 (1) ◽

Cited By ~ 13

Author(s):

Qi Zhao ◽

Xijie Wang ◽

Shuyan Wang ◽

Zheng Song ◽

Jiaxian Wang ◽

...

Keyword(s):

Stem Cells ◽

Stem Cell ◽

Embryonic Stem Cells ◽

Human Embryonic Stem Cells ◽

Pluripotent Stem Cell ◽

Embryonic Stem ◽

Induced Pluripotent Stem Cell ◽

Induced Pluripotent

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Substrates for Human Pluripotent Stem Cell Cultures in Conditioned Medium of Mesenchymal Stem Cells

Journal of Biomaterials Science Polymer Edition ◽

10.1163/092050610x545797 ◽

2012 ◽

Vol 23 (1-4) ◽

pp. 153-165 ◽

Cited By ~ 3

Author(s):

Yusuke Ueda ◽

Satoshi Fujita ◽

Tatsuya Nishigaki ◽

Yusuke Arima ◽

Hiroo Iwata

Keyword(s):

Stem Cells ◽

Mesenchymal Stem Cells ◽

Stem Cell ◽

Conditioned Medium ◽

Cell Cultures ◽

Pluripotent Stem Cell ◽

Human Pluripotent Stem Cell

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A Novel Purification Method of Murine Embryonic Stem Cell– and Human-Induced Pluripotent Stem Cell–Derived Cardiomyocytes by Simple Manual Dissociation

CrossRef Listing of Deleted DOIs ◽

10.1177/1087057111434145 ◽

2012 ◽

Vol 17 (5) ◽

pp. 683-691 ◽

Cited By ~ 13

Author(s):

Tadahiro Shinozawa ◽

Hatsue Furukawa ◽

Eimei Sato ◽

Kenji Takami

Keyword(s):

Stem Cells ◽

Stem Cell ◽

Embryonic Stem Cell ◽

Pluripotent Stem Cell ◽

Embryonic Stem ◽

Induced Pluripotent Stem Cell ◽

Drug Induced ◽

Thin Glass ◽

Murine Embryonic Stem Cell ◽

Induced Pluripotent

Cardiomyocytes derived from embryonic stem cells (ES-CMs) and induced pluripotent stem cells (iPS-CMs) are useful for toxicity and pharmacology screening. In the present study, we found that cardiomyocyte-rich beating cell clusters (CCs) emerged from murine embryonic stem cell (mESC)–derived beating EBs and from human-induced pluripotent stem cell (hiPSC)–derived beating EBs dissociated by gentle pipetting with a thin glass pipette. The percentage of cardiac troponin T (cTnT)–positive cells in the beating CCs obtained from mESC-derived and hiPSC-derived beating EBs was higher (81.5% and 91.6%, respectively) than in beating-undissociated EBs (13.7% and 67.1%, respectively). For mESCs, the yield of cTnT-positive cells from beating CCs was estimated to be 1.6 times higher than that of beating EBs. The bromodeoxyuridine labeling index of mouse ES-CMs and human iPS-CMs in beating CCs was 1.5- and 3.2-fold, respectively, greater than those in beating EBs. To investigate the utility of the cells in toxicity assessment, we showed that doxorubicin, a cardiotoxic drug, induced myofilament disruption in cardiomyocytes isolated by this method. This simple method enables preparation of mouse ES-CMs and human iPS-CMs with better proliferative activity than beating EBs not dissociated by pipetting, and the cardiomyocytes are useful for drug-induced myocardial toxicity testing.

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