Prospective identification of cardiac progenitors by a novel single cell‐based cardiomyocyte induction

Yamashita, Jun; Takano, Makoto; Hiraoka-Kanie, Mina; Shimazu, Chikashi; Yan, Peishi; Yanagi, Kentoku; Nakano, Akiko; Inoue, Emi; Kita, Fumiyo; Nishikawa, Shin‐Ichi

doi:10.1096/fj.04-3540fje

Cited by 139 publications

(168 citation statements)

References 52 publications

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Section: Embryonic Stem (Es)mentioning

confidence: 99%

“…ϩ cells may give rise to some populations of cardiomyocytes (16,17). In addition, Wnt2 expression is detected in precardiac mesoderm and derived structures such as the pericardium from as early as 7.5 days post-conception forward (24,25), and Wnt signaling is generally required for normal cardiogenesis (32,33).…”

Section: Wnt2-deficient Ebs Have Impaired Cardiomyocyte Differentiatimentioning

confidence: 99%

“…Blast colony-forming cells (BLCFCs) derived from EBs can generate hematopoietic and endothelial cell lineages, and represent the long-hypothesized hemangioblast (13,14). Flk1 ϩ cells may also give rise to smooth muscle cells (SMC) and cardiomyocytes under appropriate culture conditions (15)(16)(17).Wnts are paracrine/autocrine signaling molecules that have been implicated in many aspects of normal development. Canonical Wnt signaling acts through ␤-catenin and T-cell factor/lymphoid enhancer factor transcriptional factors that regulate target gene expression (reviewed in Refs.…”

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confidence: 99%

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Wnt2 Coordinates the Commitment of Mesoderm to Hematopoietic, Endothelial, and Cardiac Lineages in Embryoid Bodies

Wang

Gilner

Bautch

et al. 2007

Journal of Biological Chemistry

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Our recent gene expression profiling analyses demonstrated that Wnt2 is highly expressed in Flk1؉ cells, which serve as common progenitors of endothelial cells, blood cells, and mural cells. In this report, we characterize the role of Wnt2 in mesoderm development during embryonic stem (ES) cell differentiation by creating ES cell lines in which Wnt2 was deleted. Wnt2؊/؊ embryoid bodies (EBs) generated increased numbers of Flk1؉ cells and blast colony-forming cells compared with wild-type EBs, and had higher Flk1 expression at comparable stages of differentiation. Although Flk1 ؉ cells were increased, we found that endothelial cell and terminal cardiomyocyte differentiation was impaired, but hematopoietic cell differentiation was enhanced and smooth muscle cell differentiation was unchanged in Wnt2 ؊/؊ EBs. Later stage Wnt2 ؊/؊ EBs had either lower or undetectable expression of endothelial and cardiac genes compared with wild-type EBs. Consistently, vascular plexi were poorly formed and neither beating cardiomyocytes nor ␣-actinin-staining cells were detectable in later stage Wnt2 ؊/؊ EBs. In contrast, hematopoietic cell gene expression was upregulated, and the number of hematopoietic progenitor colonies was significantly enhanced in Wnt2 ؊/؊ EBs. Our data indicate that Wnt2 functions at multiple stages of development during ES cell differentiation and during the commitment and diversification of mesoderm: as a negative regulator for hemangioblast differentiation and hematopoiesis but alternatively as a positive regulator for endothelial and terminal cardiomyocyte differentiation. Embryonic stem (ES)2 cells have considerable potential for use in cellular therapies for many human diseases or disorders.However, manipulating the differentiation of pluripotent cells into a desired cell type is difficult to control. Therefore, efforts have been directed toward understanding the molecular and cellular mechanisms for maintaining ES cell pluripotency as well as for ES cell fate determination and lineage specification. Under appropriate culture conditions, ES cells will spontaneously differentiate and form embryoid bodies (EBs) that recapitulate the differentiation program of normal embryonic development. This in vitro experimental model is widely used to study mesoderm development, including the hematopoietic system, vascular endothelial cells, and cardiomyocytes (1-4).Murine hematopoietic and endothelial cell development begins in the yolk sac mesoderm at approximately day 7.5 of gestation and is marked by establishment of blood islands (5, 6). The close physical relationship between these two cell populations within blood islands has led to a widely accepted hypothesis that a common precursor for both hematopoietic and endothelial cells exists, the hemangioblast (7, 8). The receptor for vascular endothelial growth factor (VEGF), fetal liver kinase-1 (Flk1), is an early molecular marker for hemangioblasts (9 -12). Blast colony-forming cells (BLCFCs) derived from EBs can generate hematopoietic and endothelial cell lineag...

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Section: Embryonic Stem (Es)mentioning

confidence: 99%

Section: Wnt2-deficient Ebs Have Impaired Cardiomyocyte Differentiatimentioning

confidence: 99%

mentioning

confidence: 99%

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Wnt2 Coordinates the Commitment of Mesoderm to Hematopoietic, Endothelial, and Cardiac Lineages in Embryoid Bodies

Wang

Gilner

Bautch

et al. 2007

Journal of Biological Chemistry

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“…21 After 3 days of VEGFR-2 ϩ cell differentiation on OP9 cells, cultured cells were harvested and dissociated with 0.25% trypsin/EDTA (GIBCO) treatment and then placed in a 15-mL tube with medium and serum for 30 to 45 minutes at 37°C with 5% CO 2 . The cells were stained with a combination of Abs of: (1) APCconjugated podoplanin Ab and biotinylated CD31 MoAbs or VEGFR-3 goat polyclonal Abs followed by incubation with Alexa488-conjugated streptoavidin or Alexa488-conjugated antigoat Abs (Molecular Probes), or (2) VEGFR-3 goat polyclonal Abs and biotinylated CD31 MoAbs or VE-cad MoAbs followed by Alexa488-conjugated anti-goat Abs (Molecular Probes), and Alexa546-cojugated anti-rat Abs, and subjected to flow cytometry.…”

Section: Flow Cytometry and Cell Sortingmentioning

confidence: 99%

Differentiation of Lymphatic Endothelial Cells From Embryonic Stem Cells on OP9 Stromal Cells

Kono

Kubo

Shimazu

et al. 2006

ATVB

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Objectives— The discovery of vascular endothelial growth factor C (VEGF-C) and VEGF receptor-3 (VEGFR-3) has started to provide an understanding of the molecular mechanisms of lymphangiogenesis. The homeobox gene prox1 has been proven to specify lymphatic endothelial cells (ECs) from blood ECs. We investigated the process of lymphatic EC (LEC) differentiation using embryonic stem (ES) cells. Methods and Results— VEGFR-2 + cells derived from ES cells differentiated into LECs at day 3 on OP9 stromal cells defined by the expression of prox1, VEGFR-3, and another lymphatic marker podoplanin. VEGFR-2 + cells gave rise to LYVE-1 + embryonic ECs, which were negative for prox1 on day 1 but turned to prox1 + LECs by day 3. VEGFR-3-Fc or Tie2-Fc, sequestering VEGF-C or angiopoietin1 (Ang1), suppressed colony formation of LECs on OP9 cells. However, addition of VEGF-C and Ang1 in combination with VEGF to the culture of VEGFR-2 + cells on collagen-coated dishes failed to induce LECs. LEC-inducing activity of OP9 cells was fully reproduced on paraformaldehyde-fixed OP9 cells with the conditioned medium. Conclusion— We succeeded in differentiating LECs from ES cells and revealed the requirements of VEGF-C, Ang1, and other unknown factors for LEC differentiation.

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“…несущие на своих мембранах рецептор VEGFR-2), способные к дифференцировке в перициты и эндотелиоциты. После преобразования эмбриональных стволовых клеток в Flk-1 + -клетки стало возможным также получение из одной такой клетки популяции кардиомиоцитов [26,27]. Эти результаты неоднократно подтверждались другими исследователями, проводившими опыты со стволовыми клетками, которые были получены из различных источников.…”

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ROLE OF VEGFR-2 (KDR/Flk-1) IN CARDIOMYOGENESIS AND CYTOPROTECTIVE REACTIONS

Meshkov¹,

Novoselova²,

Bushmanova³

et al. 2016

СПНО (MPSE)

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Представлены современные данные о роли рецептора-2 сосудистого эндотелиального фактора роста (VEGFR2, KDR/Flk-1) в кардиомиогенезе и цитопротекторных реакциях. Показано, что функционирование рецептора VEGFR-2 на протяжении эмбриогенеза является критическим для развития организмов, поскольку нокаут гена Flk-1, кодирующего белок-рецептор VEGFR-2, на ранних стадиях эмбриогенеза приводит к гибели эмбрионов мышей из-за недостаточного образования в зародыше гемопоэтических и эндотелиальных клеток и отсутствия нормально сформированных кровеносных сосудов. На более поздних стадиях эмбриогенеза доказано участие VEGFR-2 в формировании предсердно-желудочковых клапанов сердца. Усиление регенераторного потенциала и кардиомиогенной направленности культивируемых мезенхимальных стволовых клеток достигается путем повышения в них экспрессии VEGFR-2. В экспериментах на крысах показано, что введение животным таких клеток предотвращает выраженные постинфарктные структурные изменения левого желудочка. Усиление экспрессии VEGFR-2 в кардиомиоцитах выявлено и в других моделях повреждения миокарда, в частности при моделировании антрациклиновой кардиомиопатии. Ключевые слова: миокард, кардиомиогенез, регенерация, VEGFR-2/KDR/Flk-1, клеточная терапия ROLE OF VEGFR-2 (KDR/Flk-1) IN CARDIOMYOGENESIS AND CYTOPROTECTIVE REACTIONS Meshkov I.O., Novoselova E.I., Bushmanova G.M., Semenov D.E. Institute of Molecular Pathology and Pathomorphology, Novosibirsk, e-mail: pathol@inbox.ruContemporary data on the role of vascular endothelial growth factor receptor-2 (VEGFR2, KDR/Flk-1) in cardiomyogenesis and cytoprotective reactions are presented. It is shown that the functioning of VEGFR-2 during embryogenesis is critical for the development of organisms, since the Flk-1 gene knockout in the early stages of embryogenesis leads to the death of mouse embryos due to the lack of formation of hematopoietic and endothelial cells in the fetus and the absence of normally formed blood vessels. In the later stages of embryogenesis the involvement of VEGFR-2 in the formation of the atrioventricular valves of the heart was proved. Enhancing the regenerative potential and cardiomyogenic lineage of cultured mesenchymal stem cells is done by increasing in them the expression of VEGFR-2. In experiments on rats it was shown that the injection of such cells into animals prevents post-infarction pronounced structural changes of the left ventricle. An increased expression of VEGFR-2 in cardiomyocytes also was identified in other models of myocardial damage, in particular, when modeling anthracycline cardiomyopathy.

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Prospective identification of cardiac progenitors by a novel single cell‐based cardiomyocyte induction

Cited by 139 publications

References 52 publications

Wnt2 Coordinates the Commitment of Mesoderm to Hematopoietic, Endothelial, and Cardiac Lineages in Embryoid Bodies

Wnt2 Coordinates the Commitment of Mesoderm to Hematopoietic, Endothelial, and Cardiac Lineages in Embryoid Bodies

Differentiation of Lymphatic Endothelial Cells From Embryonic Stem Cells on OP9 Stromal Cells

ROLE OF VEGFR-2 (KDR/Flk-1) IN CARDIOMYOGENESIS AND CYTOPROTECTIVE REACTIONS

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