scholarly journals The Present State and Future Perspectives of Cardiac Regenerative Therapy Using Human Pluripotent Stem Cells

2021 ◽  
Vol 8 ◽  
Author(s):  
Yusuke Soma ◽  
Yuika Morita ◽  
Yoshikazu Kishino ◽  
Hideaki Kanazawa ◽  
Keiichi Fukuda ◽  
...  
Keyword(s):  
Stem Cells ◽  
Pluripotent Stem Cells ◽  
Heart Development ◽  
Embryonic Stem ◽  
Human Pluripotent Stem Cells ◽  
Large Animal ◽  
Regenerative Therapy ◽  
Future Perspectives ◽  
Large Animal Models ◽  
Number Of Patients

The number of patients with heart failure (HF) is increasing with aging in our society worldwide. Patients with HF who are resistant to medication and device therapy are candidates for heart transplantation (HT). However, the shortage of donor hearts is a serious issue. As an alternative to HT, cardiac regenerative therapy using human pluripotent stem cells (hPSCs), such as human embryonic stem cells and induced pluripotent stem cells, is expected to be realized. Differentiation of hPSCs into cardiomyocytes (CMs) is facilitated by mimicking normal heart development. To prevent tumorigenesis after transplantation, it is important to eliminate non-CMs, including residual hPSCs, and select only CMs. Among many CM selection systems, metabolic selection based on the differences in metabolism between CMs and non-CMs is favorable in terms of cost and efficacy. Large-scale culture systems have been developed because a large number of hPSC-derived CMs (hPSC-CMs) are required for transplantation in clinical settings. In large animal models, hPSC-CMs transplanted into the myocardium improved cardiac function in a myocardial infarction model. Although post-transplantation arrhythmia and immune rejection remain problems, their mechanisms and solutions are under investigation. In this manner, the problems of cardiac regenerative therapy are being solved individually. Thus, cardiac regenerative therapy with hPSC-CMs is expected to become a safe and effective treatment for HF in the near future. In this review, we describe previous studies related to hPSC-CMs and discuss the future perspectives of cardiac regenerative therapy using hPSC-CMs.

scholarly journals Differentiation and Application of Human Pluripotent Stem Cells Derived Cardiovascular Cells for Treatment of Heart Diseases: Promises and Challenges

2021 ◽  
Vol 9 ◽  
Author(s):  
Yu Gao ◽  
Jun Pu
Keyword(s):  
Stem Cells ◽  
Pluripotent Stem Cells ◽  
Heart Diseases ◽  
Embryonic Stem ◽  
Human Pluripotent Stem Cells ◽  
Cardiac Cells ◽  
Human Embryos ◽  
Large Animal ◽  
Large Animal Models ◽  
Cardiovascular Cells

Human pluripotent stem cells (hPSCs) are derived from human embryos (human embryonic stem cells) or reprogrammed from human somatic cells (human induced pluripotent stem cells). They can differentiate into cardiovascular cells, which have great potential as exogenous cell resources for restoring cardiac structure and function in patients with heart disease or heart failure. A variety of protocols have been developed to generate and expand cardiovascular cells derived from hPSCs in vitro. Precisely and spatiotemporally activating or inhibiting various pathways in hPSCs is required to obtain cardiovascular lineages with high differentiation efficiency. In this concise review, we summarize the protocols of differentiating hPSCs into cardiovascular cells, highlight their therapeutic application for treatment of cardiac diseases in large animal models, and discuss the challenges and limitations in the use of cardiac cells generated from hPSCs for a better clinical application of hPSC-based cardiac cell therapy.

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.

scholarly journals Characterization of Endoplasmic Reticulum (ER) in Human Pluripotent Stem Cells Revealed Increased Susceptibility to Cell Death upon ER Stress

Cells ◽  
2020 ◽  
Vol 9 (5) ◽  
pp. 1078
Author(s):  
Tae Won Ha ◽  
Ji Hun Jeong ◽  
HyeonSeok Shin ◽  
Hyun Kyu Kim ◽  
Jeong Suk Im ◽  
...  
Keyword(s):  
Stem Cells ◽  
Endoplasmic Reticulum ◽  
Er Stress ◽  
Pluripotent Stem Cells ◽  
Meta Analysis ◽  
Embryonic Stem ◽  
Structural Features ◽  
Human Pluripotent Stem Cells ◽  
Differentiated Cells ◽  
Induced Apoptosis

Human pluripotent stem cells (hPSCs), such as embryonic stem cells (ESCs) and induced pluripotent stem cells (iPSCs), have a well-orchestrated program for differentiation and self-renewal. However, the structural features of unique proteostatic-maintaining mechanisms in hPSCs and their features, distinct from those of differentiated cells, in response to cellular stress remain unclear. We evaluated and compared the morphological features and stress response of hPSCs and fibroblasts. Compared to fibroblasts, electron microscopy showed simpler/fewer structures with fewer networks in the endoplasmic reticulum (ER) of hPSCs, as well as lower expression of ER-related genes according to meta-analysis. As hPSCs contain low levels of binding immunoglobulin protein (BiP), an ER chaperone, thapsigargin treatment sharply increased the gene expression of the unfolded protein response. Thus, hPSCs with decreased chaperone function reacted sensitively to ER stress and entered apoptosis faster than fibroblasts. Such ER stress-induced apoptotic processes were abolished by tauroursodeoxycholic acid, an ER-stress reliever. Hence, our results revealed that as PSCs have an underdeveloped structure and express fewer BiP chaperone proteins than somatic cells, they are more susceptible to ER stress-induced apoptosis in response to stress.

scholarly journals Establishing a deeper understanding of the osteogenic differentiation of monolayer cultured human pluripotent stem cells using novel and detailed analyses

2021 ◽  
Vol 12 (1) ◽  
Author(s):  
Ping Zhou ◽  
Jia-Min Shi ◽  
Jing-E Song ◽  
Yu Han ◽  
Hong-Jiao Li ◽  
...  
Keyword(s):  
Stem Cells ◽  
Osteogenic Differentiation ◽  
Pluripotent Stem Cells ◽  
Embryonic Stem ◽  
Telomerase Activity ◽  
Human Pluripotent Stem Cells ◽  
Cell Counting ◽  
Cell Type ◽  
Alizarin Red

Abstract Background Derivation of osteoblast-like cells from human pluripotent stem cells (hPSCs) is a popular topic in bone tissue engineering. Although many improvements have been achieved, the low induction efficiency because of spontaneous differentiation hampers their applications. To solve this problem, a detailed understanding of the osteogenic differentiation process of hPSCs is urgently needed. Methods Monolayer cultured human embryonic stem cells and human-induced pluripotent stem cells were differentiated in commonly applied serum-containing osteogenic medium for 35 days. In addition to traditional assays such as cell viability detection, reverse transcription-polymerase chain reaction, immunofluorescence, and alizarin red staining, we also applied studies of cell counting, cell telomerase activity, and flow cytometry as essential indicators to analyse the cell type changes in each week. Results The population of differentiated cells was quite heterogeneous throughout the 35 days of induction. Then, cell telomerase activity and cell cycle analyses have value in evaluating the cell type and tumourigenicity of the obtained cells. Finally, a dynamic map was made to integrate the analysis of these results during osteogenic differentiation of hPSCs, and the cell types at defined stages were concluded. Conclusions Our results lay the foundation to improve the in vitro osteogenic differentiation efficiency of hPSCs by supplementing with functional compounds at the desired stage, and then establishing a stepwise induction system in the future.

scholarly journals Probing the mechanism for hydrogel-based stasis induction in human pluripotent stem cells: is the chemical functionality of the hydrogel important?

2020 ◽  
Vol 11 (1) ◽  
pp. 232-240 ◽  
Author(s):  
M. Sponchioni ◽  
C. T. O'Brien ◽  
C. Borchers ◽  
E. Wang ◽  
M. N. Rivolta ◽  
...  
Keyword(s):  
Stem Cells ◽  
Stem Cell ◽  
Block Copolymer ◽  
Embryonic Stem Cell ◽  
Pluripotent Stem Cells ◽  
Human Embryonic Stem Cell ◽  
Embryonic Stem ◽  
Human Pluripotent Stem Cells ◽  
Essential Parameter ◽  
Chemical Functionality

It is shown that hydroxyl functionality is required to induce stasis in human embryonic stem cell colonies immersed within wholly synthetic block copolymer worm gels with comparable storage moduli. Thus gel softness does not appear to be an essential parameter for stasis induction.

scholarly journals An improved method to produce clinical scale natural killer cells from human pluripotent stem cells

2019 ◽  
Author(s):  
Huang Zhu ◽  
Dan S. Kaufman
Keyword(s):  
Stem Cells ◽  
Nk Cells ◽  
Natural Killer ◽  
Pluripotent Stem Cells ◽  
Nk Cell ◽  
Embryonic Stem ◽  
Human Pluripotent Stem Cells ◽  
Genetically Engineered ◽  
Cell Populations ◽  
Clinical Scale

AbstractHuman natural killer (NK) cell-based adoptive anti-cancer immunotherapy has gained intense interest with many clinical trials actively recruiting patients to treat a variety of both hematological malignancies and solid tumors. Most of these trials use primary NK cells isolated either from peripheral blood (PB-NK cells) or umbilical cord blood (UCB-NK cells), though these sources require NK cell collection for each patient leading to donor variability and heterogeneity in the NK cell populations. In contrast, NK cells derived human embryonic stem cells (hESC-NK cells) or induced pluripotent stem cells (hiPSC-NK cells) provide more homogeneous cell populations that can be grown at clinical scale, and genetically engineered if desired. These characteristics make hESC/iPSC-derived NK cells an ideal cell population for developing standardized, “off-the-shelf” immunotherapy products. Additionally, production of NK cells from undifferentiated human pluripotent stem cells enables studies to better define pathways that regulate human NK cell development and function. Our group previously established a stromal-free, two-stage culture system to derive NK cells from hESC/hiPSC in vitro followed by clinical-scale expansion of these cells using interleukin-21 expressing artificial antigen-presenting cells. However, prior to differentiation, this method requires single cell adaption of hESCs/hiPSCs which takes months. Recently we optimized this method by adapting the mouse embryonic fibroblast-dependent hESC/hiPSC to feeder-free culture conditions. These feeder-free hESC/hiPSCs are directly used to generate hemato-endothelial precursor cells. This new method produces mature, functional NK cells with higher efficiency to enable rapid production of an essentially unlimited number of homogenous NK cells that can be used for standardized, targeted immunotherapy for the treatment of refractory cancers and infectious diseases.

scholarly journals Generation of CD34+CD43+ hematopoietic progenitors to induce thymocytes from human pluripotent stem cells

2021 ◽  
Author(s):  
Lea Flippe ◽  
Anne Gaignerie ◽  
Celine Serazin ◽  
Olivier Baron ◽  
Xavier Saulquin ◽  
...  
Keyword(s):  
Stem Cells ◽  
T Cells ◽  
Pluripotent Stem Cells ◽  
High Efficiency ◽  
Embryonic Stem ◽  
Human Pluripotent Stem Cells ◽  
Blood Type ◽  
Hematopoietic Stem ◽  
Similar Profile

Immunotherapy using primary T cells has revolutionized medical care in some pathologies in recent years but limitations associated to challenging cell genome edition, insufficient cell number production, the use of only autologous cells and lack of product standardization have limited its uses in the clinic. The alternative use of T cells generated in vitro from human pluripotent stem cells (hPSCs) offers great advantages by providing a self-renewing source of T cells that can be readily genetically modified and facilitate the use of standardized universal off-the-shelf allogeneic cell products and rapid clinic access. However, despite their potential, the feasibility and functionality of T-cells differentiated from hPSCs needs better comprehension before moving to the clinic. In this study, we generated human induced pluripotent stem cells from T-cells (T-iPSCs) allowing preservation of already recombined TCR, with the same properties as human embryonic stem cells (hESCs). Based on these cells, we differentiated with high efficiency hematopoietic progenitor stem cells (HPSCs), capable of self-renewal and differentiation into any cell blood type, and then DN3a thymic progenitors from several T-iPSC lines. To better comprehend differentiation, we analyzed the transcriptomic profiles of the different cell types and demonstrated that HPSCs differentiated from hiPSCs had a very similar profile to cord blood hematopoietic stem cells (HSCs). Furthermore, differentiated T-cell progenitors had a similar profile to thymocytes at the DN3a stage of thymic lymphopoiesis. Therefore, with this approach, we were able to regenerate precursors of therapeutic human T cells to potentially treat a wide number of diseases.

Abstract 227: Cardiomyocytes from Human Pluripotent Stem Cells Predict Chemotherapy-Induced Cardiotoxicity

2012 ◽  
Vol 111 (suppl_1) ◽  
Author(s):  
Maxime Mioulane ◽  
Gabor Foldes ◽  
Sian Harding
Keyword(s):  
Stem Cells ◽  
Cell Death ◽  
Pluripotent Stem Cells ◽  
Fluorescent Dyes ◽  
Embryonic Stem ◽  
Chemotherapeutic Agents ◽  
Human Pluripotent Stem Cells ◽  
Toxicity Tests ◽  
Dependent Manner ◽  
Isolated Cells

The potential of human pluripotent stem cells-derived cardiomyocytes (hPSC-CM) to mimic human cardiac toxicity tests is largely unknown. Here we investigate the interaction between the chemotherapeutic agents Doxorubicin (DOX) and Lapatinib (LAP) and the protective effect of beta2 adrenergic signalling in hPSC-CM and murine myocytes. We hypothesized that death and survival pathways in hPSC-CM differ from those in animal-derived myocytes and thereby may predict better clinical outcome of chemotherapeutics. Human induced pluripotent or embryonic stem cell H7 cell lines were differentiated into cardiomyocytes (hiPSC-CM and hESC-CM, respectively), treated with drugs and stained with fluorescent dyes (Hoechst 33342, mitochondrial probes, effector caspases activation and impermeant nuclear dyes). Cell death profile was generated from high content screening and analysis (Cellomics). In isolated hPSC-CM, the anthracycline DOX induced apoptosis in a concentration-dependent manner (1-100µM), whereas LAP did not produce any toxicity. However, the combination of the two chemotherapeutic agents worsened the toxicity profile compared to DOX alone (p<0.001, n=12), consistent with clinical observations. In contrast, rat neonates were sensitive to LAP and showed a different cell death profile in DOX and drug combination experiments. Activation of apoptotic caspases in hESC-CM clusters was coincident with the dissipation of the mitochondrial membrane potential, while the potential was preserved in isolated cells during this phase. Beta2 adrenergic stimulation did not reverse DOX pro-apoptotic effect but modified toxicity-related nuclear events. In conclusion, we have shown that hPSC-CM reproduce the clinical adverse cardiac effects after treatment with anti-cancer drugs, making these cells a more relevant system for future drug development than animal-derived cells. However, our data suggest that cell death pathways involved may depend on culture condition for cardiomyocytes.

Evidence That Human Very Small Embryonic-Like Stem Cells (VSELs) Are Mobilized by G-CSF Into Peripheral Blood: A Novel Strategy to Obtain Human Pluripotent Stem Cells for Regenerative Medicine.

Blood ◽  
2009 ◽  
Vol 114 (22) ◽  
pp. 1474-1474
Author(s):  
Satish Medicetty ◽  
Mariusz Z Ratajczak ◽  
Magdalena J Kucia ◽  
Ewa K. Zuba-Surma ◽  
Izabela Klich ◽  
...  
Keyword(s):  
Stem Cells ◽  
Regenerative Medicine ◽  
Pluripotent Stem Cells ◽  
Peripheral Blood ◽  
Embryonic Stem ◽  
Human Pluripotent Stem Cells ◽  
Employment Equity ◽  
Hematopoietic Stem ◽  
Equity Ownership ◽  
Novel Strategy

Abstract Abstract 1474 Poster Board I-497 We previously demonstrated that human cord blood contains a population of small (smaller in size than erythrocytes) CXCR4+CD133+CD34+SSEA-4+Oct-4+lin−CD45− cells (Leukemia 2007:21;297-303) and that these cells are mobilized into peripheral blood during tissue organ damage as seen for example in heart infarct (J. Am. Coll. Cardiol., 2009:53;1-9.) or stroke (Stroke. 2009:40;1237.). Similar cells were also reported in murine organs, and more importantly we described that these cells may differentiate in vitro into cells from all three germ layers (Leukemia 2006:20;857–869). To explore the possibility that human VSELs could become a source of pluripotent stem cells in regenerative medicine, our goal was to develop an efficient strategy to isolate these cells from adult patients. To test if VSELs similarly to their murine counterparts could be mobilized into peripheral blood after granulocyte colony stimulating factor (G-CSF) injection (Stem Cells 2008:26;2083-2092), we enrolled a group of young healthy donors who were mobilized for two consecutive days using G-CSF (480 μg/day subcutaneously). On the third day nucleated cells (TNC) were collected by apheresis. We evaluated number of VSELs in peripheral blood (PB) samples before and after G-CSF mobilization as well as the final number in the apheresis product. At least 1 million of TNC were acquired and analyzed by FACS Diva software. Three different fractions of non-hematopoietic stem cells enriched for VSELs (Lin−/CD45−/CD133+, Lin−/CD45−/CD34+, Lin−/CD45−/CXCR4+) as well as their CD45 positive hematopoietic counterparts were analyzed. The absolute numbers of cells from each population, contained in 1 μL of sample, were computed based on percent content of each population and TNC count for each individual sample. Results show that after G-CSF mobilization, human peripheral blood contains a population of lin− CD45− mononuclear cells that express CXCR4, CD34 and CD133 antigens. These lin− CD45− CXCR4+ CD133+ CD34+ cells are highly enriched for mRNA for intra-nuclear pluripotent embryonic transcription factors such as Oct-4, Sox2 and Nanog. More importantly we found that Oct-4 was expressed in nuclei of mobilized VSELs and that these cells also express the cell surface marker SSEA-4, the early embryonic glycolipid antigen commonly used as a marker for undifferentiated pluripotent human embryonic stem cells. We observed that these adult peripheral blood-derived VSELs are slightly larger than their counterparts identified in adult murine bone marrow, but are still very small. In addition, they also possess large nuclei containing embryonic-type unorganized euchromatin. Before G-CSF mobilization only very few VSELs were detectable in peripheral blood, whereas following G-CSF induced mobilization there was a very significant increase with in excess of 106 VSELs present in the apheresis product representing less than 0.01% of TNC. We postulate that while VSELs are relatively rare cells, they are mobilized into peripheral blood and that G-CSF induced mobilization could become a novel strategy to obtain human pluripotent stem cells for regenerative medicine. Disclosures: Medicetty: NeoStem Inc: Employment, Equity Ownership. Marasco: NeoStem Inc: Consultancy, Equity Ownership, Membership on an entity's Board of Directors or advisory committees. Rodgerson: NeoStem Inc: Employment, Equity Ownership.

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