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  • 1
    Online Resource
    Online Resource
    Human embryonic stem cell–derived cardiomyocytes can mobilize 1,4,5‐inositol trisphosphate–operated [Ca 2+ ] i stores : The functionality of angiotensin‐II/endothelin‐1 signaling pathways
    Wiley ; 2010
    In:  Annals of the New York Academy of Sciences Vol. 1188, No. 1 ( 2010-02), p. 68-77
    Details
    In: Annals of the New York Academy of Sciences, Wiley, Vol. 1188, No. 1 ( 2010-02), p. 68-77
    Abstract: Because previous findings showed that in human embryonic stem cell–derived cardiomyocytes (hESC‐CM) the machinery for Ca 2+ ‐induced release of calcium is immature, we tested the hypothesis that hESC‐CM contain functional 1,4,5‐inositol triphosphate (IP 3 )–operated intracellular Ca 2+ ([Ca 2+ ] i ) stores. We investigated the effects of angiotensin II (AT‐II) and endothelin 1 (ET‐1), which activate the 1,4,5‐IP 3 pathway, on [Ca 2+ ] i transients and contractions in hESC‐CM. Our major findings were that in hESC‐CM, both AT‐II (10 −9 –10 −7 M) and ET‐1 (10 −9 –10 −7 M) exert inotropic and lusitropic effects. The involvement of 1,4,5‐IP 3 ‐dependent intracellular Ca 2+ release in AT‐I–induced effects was supported by these findings: the effects of AT‐II were blocked by 2‐aminoethoxyphenyl borate (2‐APB, a 1,4,5‐IP 3 receptor blocker) and U73122 (a phosopholipase C blocker); and hESC‐CM express AT‐II type 1 and IP 3 type I and II receptors as determined by fluorescence immunostaining. In conclusion, hESC‐CM exhibit functional AT‐II and ET‐1 signaling pathways, as well as 1,4,5‐IP 3 –operated releasable Ca 2+ stores.
    Type of Medium: Online Resource
    ISSN: 0077-8923 , 1749-6632
    URL: Issue
    URL: Article
    DOI: 10.1111/nyas.2010.1188.issue-1
    DOI: 10.1111/j.1749-6632.2009.05085.x
    RVK:
    TD 7650
    Language: English
    Publisher: Wiley
    Publication Date: 2010
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  • 2
    Online Resource
    Online Resource
    Functional Properties of Human Embryonic Stem Cell‐Derived Cardiomyocytes
    Wiley ; 2005
    In:  Annals of the New York Academy of Sciences Vol. 1047, No. 1 ( 2005-06), p. 66-75
    Details
    In: Annals of the New York Academy of Sciences, Wiley, Vol. 1047, No. 1 ( 2005-06), p. 66-75
    Abstract: A bstract : Regeneration of the diseased myocardium by cardiac cell transplantation is an attractive therapeutic modality. Yet, because the transplanted cardiomyocytes should functionally integrate within the diseased myocardium, it is preferable that their properties resemble those of the host. To determine the functional adaptability of human embryonic stem cell‐derived cardiomyocytes (hESC‐CM) to the host myocardium, the authors investigated the excitation‐contraction (E‐C) coupling and the responsiveness to common physiological stimuli. The main findings are: (1) hESC‐CM readily respond to electrical pacing and generate corresponding [Ca 2+ ] i transients (measured by fura‐2 fluorescence) and contractions (measured by video edge detector). (2) In contrast to the mature myocardium, hESC‐CM display negative force‐frequency relations. (3) The hESC‐CM contraction is dependent on [Ca 2+ ] o and blocked by verapamil. (4) Surprisingly, ryanodine, the sarcoplasmic‐endoplasmic reticulum Ca 2+ ‐ATPase inhibitor thapsigargin, and caffeine do not affect the [Ca 2+ ] i transient or contraction. Collectively, these results indicate that at the developmental stage of 45 to 60 days, the contraction is largely dependent on [Ca 2+ ] o rather than on sarcoplasmic reticulum (SR) Ca 2+ stores. The results show for the first time that the E‐C coupling properties of hESC‐CM differ from the adult myocardium, probably due to immature SR function. Based on these findings, genetic manipulation of hESC‐CM toward the adult myocardium should be considered.
    Type of Medium: Online Resource
    ISSN: 0077-8923 , 1749-6632
    URL: Article
    DOI: 10.1196/annals.1341.006
    RVK:
    TD 7650
    Language: English
    Publisher: Wiley
    Publication Date: 2005
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    detail.hit.zdb_id: 211003-9
    detail.hit.zdb_id: 2071584-5
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  • 3
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    Characterization of the expression of MHC proteins in human embryonic stem cells
    Proceedings of the National Academy of Sciences ; 2002
    In:  Proceedings of the National Academy of Sciences Vol. 99, No. 15 ( 2002-07-23), p. 9864-9869
    Details
    In: Proceedings of the National Academy of Sciences, Proceedings of the National Academy of Sciences, Vol. 99, No. 15 ( 2002-07-23), p. 9864-9869
    Abstract: Human embryonic stem (ES) cells are pluripotent cells that may be used in transplantation medicine. These cells can be induced to differentiate into cells from the three embryonic germ layers both in vivo and in vitro . To determine whether human ES cells might be rejected after transplantation, we examined cell surface expression of the MHC proteins in these cells. Our results show very low expression levels of MHC class I (MHC-I) proteins on the surface of human ES cells that moderately increase on in vitro or in vivo differentiation. A dramatic induction of MHC-I proteins was observed when the cells were treated with IFN-γ but not with IFN-α or -β. However, all three IFNs induced expression of MHC-I proteins in differentiated human ES cells. MHC-II proteins and HLA-G were not expressed on the surface of undifferentiated or differentiated cells. Ligands for natural killer cell receptors were either absent or expressed in very low levels in human ES cells and in their differentiated derivatives. In accordance, natural killer cytotoxic assays demonstrated only limited lysis of both undifferentiated and differentiated cells. To initiate a histocompatibility databank of human ES cells, we have isotyped several of the published ES cell lines for their human leukocyte antigens. In conclusion, our results demonstrate that human ES cells can express high levels of MHC-I proteins and thus may be rejected on transplantation.
    Type of Medium: Online Resource
    ISSN: 0027-8424 , 1091-6490
    URL: Article
    DOI: 10.1073/pnas.142298299
    RVK:
    TA 1000
    RVK:
    WA 15000
    Language: English
    Publisher: Proceedings of the National Academy of Sciences
    Publication Date: 2002
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  • 4
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    Online Resource
    Differentiation of human embryonic stem cells on three-dimensional polymer scaffolds
    Proceedings of the National Academy of Sciences ; 2003
    In:  Proceedings of the National Academy of Sciences Vol. 100, No. 22 ( 2003-10-28), p. 12741-12746
    Details
    In: Proceedings of the National Academy of Sciences, Proceedings of the National Academy of Sciences, Vol. 100, No. 22 ( 2003-10-28), p. 12741-12746
    Abstract: Human embryonic stem (hES) cells hold promise as an unlimited source of cells for transplantation therapies. However, control of their proliferation and differentiation into complex, viable 3D tissues is challenging. Here we examine the use of biodegradable polymer scaffolds for promoting hES cell growth and differentiation and formation of 3D structures. We show that complex structures with features of various committed embryonic tissues can be generated, in vitro , by using early differentiating hES cells and further inducing their differentiation in a supportive 3D environment such as poly(lactic- co -glycolic acid)/poly( l -lactic acid) polymer scaffolds. We found that hES cell differentiation and organization can be influenced by the scaffold and directed by growth factors such as retinoic acid, transforming growth factor β, activin-A, or insulin-like growth factor. These growth factors induced differentiation into 3D structures with characteristics of developing neural tissues, cartilage, or liver, respectively. In addition, formation of a 3D vessel-like network was observed. When transplanted into severe combined immunodeficient mice, the constructs continue to express specific human proteins in defined differentiated structures and appear to recruit and anastamose with the host vasculature. This approach provides a unique culture system for addressing questions in cell and developmental biology, and provides a potential mechanism for creating viable human tissue structures for therapeutic applications.
    Type of Medium: Online Resource
    ISSN: 0027-8424 , 1091-6490
    URL: Article
    DOI: 10.1073/pnas.1735463100
    RVK:
    TA 1000
    RVK:
    WA 15000
    Language: English
    Publisher: Proceedings of the National Academy of Sciences
    Publication Date: 2003
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  • 5
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    SK4 Ca 2+ activated K + channel is a critical player in cardiac pacemaker derived from human embryonic stem cells
    Proceedings of the National Academy of Sciences ; 2013
    In:  Proceedings of the National Academy of Sciences Vol. 110, No. 18 ( 2013-04-30)
    Details
    In: Proceedings of the National Academy of Sciences, Proceedings of the National Academy of Sciences, Vol. 110, No. 18 ( 2013-04-30)
    Abstract: Proper expression and function of the cardiac pacemaker is a critical feature of heart physiology. Two main mechanisms have been proposed: ( i ) the “voltage-clock,” where the hyperpolarization-activated funny current I f causes diastolic depolarization that triggers action potential cycling; and ( ii ) the “Ca 2+ clock,” where cyclical release of Ca 2+ from Ca 2+ stores depolarizes the membrane during diastole via activation of the Na + –Ca 2+ exchanger. Nonetheless, these mechanisms remain controversial. Here, we used human embryonic stem cell-derived cardiomyocytes (hESC-CMs) to study their autonomous beating mechanisms. Combined current- and voltage-clamp recordings from the same cell showed the so-called “voltage and Ca 2+ clock” pacemaker mechanisms to operate in a mutually exclusive fashion in different cell populations, but also to coexist in other cells. Blocking the “voltage or Ca 2+ clock” produced a similar depolarization of the maximal diastolic potential (MDP) that culminated by cessation of action potentials, suggesting that they converge to a common pacemaker component. Using patch-clamp recording, real-time PCR, Western blotting, and immunocytochemistry, we identified a previously unrecognized Ca 2+ -activated intermediate K + conductance (IK Ca , KCa3.1, or SK4) in young and old stage-derived hESC-CMs. IK Ca inhibition produced MDP depolarization and pacemaker suppression. By shaping the MDP driving force and exquisitely balancing inward currents during diastolic depolarization, IK Ca appears to play a crucial role in human embryonic cardiac automaticity.
    Type of Medium: Online Resource
    ISSN: 0027-8424 , 1091-6490
    URL: Article
    DOI: 10.1073/pnas.1221022110
    RVK:
    TA 1000
    RVK:
    WA 15000
    Language: English
    Publisher: Proceedings of the National Academy of Sciences
    Publication Date: 2013
    detail.hit.zdb_id: 209104-5
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  • 6
    Online Resource
    Online Resource
    The Ovarian Prorenin‐Angiotensin System a : Lessons from IVF
    Wiley ; 1988
    In:  Annals of the New York Academy of Sciences Vol. 541, No. 1 ( 1988-10), p. 179-189
    Details
    In: Annals of the New York Academy of Sciences, Wiley, Vol. 541, No. 1 ( 1988-10), p. 179-189
    Type of Medium: Online Resource
    ISSN: 0077-8923 , 1749-6632
    URL: Issue
    URL: Article
    DOI: 10.1111/nyas.1988.541.issue-1
    DOI: 10.1111/j.1749-6632.1988.tb22254.x
    RVK:
    TD 7650
    Language: English
    Publisher: Wiley
    Publication Date: 1988
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  • 7
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    Endothelial cells derived from human embryonic stem cells
    Proceedings of the National Academy of Sciences ; 2002
    In:  Proceedings of the National Academy of Sciences Vol. 99, No. 7 ( 2002-04-02), p. 4391-4396
    Details
    In: Proceedings of the National Academy of Sciences, Proceedings of the National Academy of Sciences, Vol. 99, No. 7 ( 2002-04-02), p. 4391-4396
    Abstract: Human embryonic stem cells have the potential to differentiate into various cell types and, thus, may be useful as a source of cells for transplantation or tissue engineering. We describe here the differentiation steps of human embryonic stem cells into endothelial cells forming vascular-like structures. The human embryonic-derived endothelial cells were isolated by using platelet endothelial cell-adhesion molecule-1 (PECAM1) antibodies, their behavior was characterized in vitro and in vivo , and their potential in tissue engineering was examined. We show that the isolated embryonic PECAM1+ cells, grown in culture, display characteristics similar to vessel endothelium. The cells express endothelial cell markers in a pattern similar to human umbilical vein endothelial cells, their junctions are correctly organized, and they have high metabolism of acetylated low-density lipoprotein. In addition, the cells are able to differentiate and form tube-like structures when cultured on matrigel. In vivo , when transplanted into SCID mice, the cells appeared to form microvessels containing mouse blood cells. With further studies, these cells could provide a source of human endothelial cells that could be beneficial for potential applications such as engineering new blood vessels, endothelial cell transplantation into the heart for myocardial regeneration, and induction of angiogenesis for treatment of regional ischemia.
    Type of Medium: Online Resource
    ISSN: 0027-8424 , 1091-6490
    URL: Article
    DOI: 10.1073/pnas.032074999
    RVK:
    TA 1000
    RVK:
    WA 15000
    Language: English
    Publisher: Proceedings of the National Academy of Sciences
    Publication Date: 2002
    detail.hit.zdb_id: 209104-5
    detail.hit.zdb_id: 1461794-8
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    SSG: 12
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  • 8
    Online Resource
    Online Resource
    Effects of eight growth factors on the differentiation of cells derived from human embryonic stem cells
    Proceedings of the National Academy of Sciences ; 2000
    In:  Proceedings of the National Academy of Sciences Vol. 97, No. 21 ( 2000-10-10), p. 11307-11312
    Details
    In: Proceedings of the National Academy of Sciences, Proceedings of the National Academy of Sciences, Vol. 97, No. 21 ( 2000-10-10), p. 11307-11312
    Abstract: Human embryonic stem (ES) cells are pluripotent cells derived from the inner cell mass of in vitro fertilized human blastocysts. We examined the potential of eight growth factors [basic fibroblast growth factor (bFGF), transforming growth factor β1 (TGF-β1), activin-A, bone morphogenic protein 4 (BMP-4), hepatocyte growth factor (HGF), epidermal growth factor (EGF), β nerve growth factor (βNGF), and retinoic acid] to direct the differentiation of human ES-derived cells in vitro . We show that human ES cells that have initiated development as aggregates (embryoid bodies) express a receptor for each of these factors, and that their effects are evident by differentiation into cells with different epithelial or mesenchymal morphologies. Differentiation of the cells was assayed by expression of 24 cell-specific molecular markers that cover all embryonic germ layers and 11 different tissues. Each growth factor has a unique effect that may result from directed differentiation and/or cell selection, and we can divide the overall effects of the factors into three categories: growth factors (Activin-A and TGFβ1) that mainly induce mesodermal cells; factors (retinoic acid, EGF, BMP-4, and bFGF) that activate ectodermal and mesodermal markers; and factors (NGF and HGF) that allow differentiation into the three embryonic germ layers, including endoderm. None of the growth factors directs differentiation exclusively to one cell type. This analysis sets the stage for directing differentiation of human ES cells in culture and indicates that multiple human cell types may be enriched in vitro by specific factors.
    Type of Medium: Online Resource
    ISSN: 0027-8424 , 1091-6490
    URL: Article
    DOI: 10.1073/pnas.97.21.11307
    RVK:
    TA 1000
    RVK:
    WA 15000
    Language: English
    Publisher: Proceedings of the National Academy of Sciences
    Publication Date: 2000
    detail.hit.zdb_id: 209104-5
    detail.hit.zdb_id: 1461794-8
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  • 9
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    Embryonic Stem Cell Lines Derived from Human Blastocysts
    American Association for the Advancement of Science (AAAS) ; 1998
    In:  Science Vol. 282, No. 5391 ( 1998-11-06), p. 1145-1147
    Details
    In: Science, American Association for the Advancement of Science (AAAS), Vol. 282, No. 5391 ( 1998-11-06), p. 1145-1147
    Abstract: Human blastocyst-derived, pluripotent cell lines are described that have normal karyotypes, express high levels of telomerase activity, and express cell surface markers that characterize primate embryonic stem cells but do not characterize other early lineages. After undifferentiated proliferation in vitro for 4 to 5 months, these cells still maintained the developmental potential to form trophoblast and derivatives of all three embryonic germ layers, including gut epithelium (endoderm); cartilage, bone, smooth muscle, and striated muscle (mesoderm); and neural epithelium, embryonic ganglia, and stratified squamous epithelium (ectoderm). These cell lines should be useful in human developmental biology, drug discovery, and transplantation medicine.
    Type of Medium: Online Resource
    ISSN: 0036-8075 , 1095-9203
    URL: Article
    DOI: 10.1126/science.282.5391.1145
    RVK:
    TA 1000
    RVK:
    WA 15000
    Language: English
    Publisher: American Association for the Advancement of Science (AAAS)
    Publication Date: 1998
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    detail.hit.zdb_id: 2066996-3
    detail.hit.zdb_id: 2060783-0
    SSG: 11
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