Coculture of Embryonic Ventricular Myocytes and Mouse Embryonic Stem Cell Enhance Intercellular Signaling by Upregulation of Connexin43

Tang, Ming; Liang, Huamin; Li, Yuting; Wang, Jian; Song, Yuanlong; Zheng, Yongqiang; Xi, Jun; Zhang, Jinxia; Hescheler, Jürgen; Zhu, Minyan

doi:10.1159/000350124

Cited by 6 publications

(4 citation statements)

References 33 publications

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“…A regenerative potential of the CD117 + SC population might be attributed to their differentiation into an endothelial-like phenotype, as shown in a 2D-Matrigel assay and with patch-clamp measurements in recent studies published by our group [17]. Using a co-culture setting involving the well characterized HL-1 cell line, we could show proof-of-principle that cellto-cell contact is a pre-requisite for CD117 + SC to improve cardiomyocyte vitality [38,39]. Therefore, our findings are in line with a recent study demonstrating that cell-to-cell contact induces anti-apoptotic pathways in hematopoietic stem cells (HSC) indicating cardioprotective effects of HSC on cardiomyocytes in a co-culture of HSC with neonatal rat ventricular myocytes [40].…”

Section: Discussionsupporting

confidence: 58%

Exploiting AT2R to Improve CD117 Stem Cell Function In Vitro and In Vivo - Perspectives for Cardiac Stem Cell Therapy

Ludwig¹,

Tölk²,

Skorska³

et al. 2015

Cell Physiol Biochem

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Background/Aims: CD117⁺ stem cell (SC) based therapy is considered an alternative therapeutic option for terminal heart disease. However, controversies exist on the effects of CD117⁺ SC implantation. In particular, the link between CD117⁺ SC function and angiotensin-II-type-2 receptor (AT2R) after MI is continuously discussed. We therefore asked whether 1) AT2R stimulation influences CD117⁺ SC properties in vitro and, 2) which effects can be ascribed to AT2R stimulation in vivo. Methods: We approached AT2R stimulation with Angiotensin II while simultaneously blocking its opponent receptor AT1 with Losartan. CD117 effects were dissected using a 2D-Matrigel assay and HL-1 co-culture in vitro. A model of myocardial infarction, in which we implanted EGFP+ CD117 SC, was further applied. Results: While we found indications for AT2R driven vasculogenesis in vitro, co-culture experiments revealed that CD117⁺ SC improve vitality of cardiomyocytes independently of AT2R function. Likewise, untreated CD117⁺ SC had a positive effect on cardiac function and acted cardioprotective in vivo. Conclusions: Therefore, our data show that transient AT2R stimulation does not significantly add to the beneficial actions of CD117⁺ SC in vivo. Yet, exploiting AT2R driven vasculogenis via an optimized AT2R stimulation protocol may become a promising tool for cardiac SC therapy.

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Section: Discussionsupporting

confidence: 58%

Exploiting AT2R to Improve CD117 Stem Cell Function In Vitro and In Vivo - Perspectives for Cardiac Stem Cell Therapy

Ludwig¹,

Tölk²,

Skorska³

et al. 2015

Cell Physiol Biochem

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“…Previously, many reports suggest that co-cultured stem cells with other cell line results in significantly altered phenotype and differentiated characteristics [ 16 – 18 ]. Here, co-culture of EBs and NCMs was established in a transwell insert system, while EBs grown alone as controls.…”

Section: Discussionmentioning

confidence: 99%

Co-culture with neonatal cardiomyocytes enhances the proliferation of iPSC-derived cardiomyocytes via FAK/JNK signaling

Wang

Huang

et al. 2016

BMC Dev Biol

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BackgroundWe previously reported that the pluripotent stem cells can differentiate into cardiomyocytes (CMs) by co-culture with neonatal CMs (NCMs) in vitro. However, the involving mechanism is not clear.MethodsMouse induced pluripotent stem cells (iPSCs) were cultured in hanging drops to form embryoid bodies (EBs) and to induce myocardial differentiation. Co-culture of EBs and NCMs was established in a transwell insert system, while EBs grown alone in the wells were used as controls.ResultsCo-culture with NCMs markedly increased the generation of functional CMs from iPSCs. The focal adhesion kinase (FAK) phosphorylation, and c-Jun N-terminal kinase (JNK) phosphorylation in co-culture were higher than that in EBs grown alone. Treating FAK small interfering RNA (FAK siRNA) or specific inhibitor for JNK (SP600125) to iPSCs significantly reduced the phosphorylation of JNK and the expressions of Mef2c and Bcl-2. The expressions of cTnT and MLC-2V were also decreased. Our results revealed that co-culture with NCMs significantly enhance the differentiation ability of iPSCs by increasing Mef2c and Bcl-2 expressions concomitantly with a marked augment on cell proliferation through JNK signaling pathways.ConclusionsThese findings indicated that co-culture of EBs with NCMs induces genes expressed in a mature pattern and stimulates the proliferation of iPSC-derived CMs (iPS-CMs) by activating FAK/JNK signaling.Electronic supplementary materialThe online version of this article (doi:10.1186/s12861-016-0112-2) contains supplementary material, which is available to authorized users.

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“…The channel permits the intercellular passage of small molecules (generally less than 1000 Da), such as inorganic ions (e.g., Na+ and K+), second messengers (e.g., Ca 2+ , IP3, and cAMP), hormones, and other metabolites, enabling the neighboring cells to achieve gap junction intercellular communication (GJIC). Connexins and GJIC play crucial roles in the regulation of embryonic development, maintenance of tissue/organ homeostasis, cellular metabolism, morphogenesis, and growth control [14][15][16]. Of note, Cx43 is the most abundant connexin, and it is the most closely related to cell growth.…”

Section: Introductionmentioning

confidence: 99%

Beyond Gap Junction Channel Function: the Expression of Cx43 Contributes to Aldosterone-Induced Mesangial Cell Proliferation via the ERK1/2 and PKC Pathways

Zhang

Han

Wang³

et al. 2015

Cell Physiol Biochem

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Aims: This study aimed to explore the precise mechanism and signaling pathways of mesangial cell (MC) proliferation from a new point of view considering Connexin 43 (Cx43). Methods: MC proliferation was measured by the incorporation of 3H-thymidine (3H-TdR). Cx43 was over-expressed in MC cells using lipofectamine 2000, and the expression level was tested with reverse transcription-polymerase chain reaction (RT-PCR) and Western blot analyses. The gap junction channel function was explored by Lucifer Yellow scrape loading and dye transfer (SLDT), and the intracellular calcium concentrations ([Ca²⁺]_i) were characterized by confocal microscopy on cells loaded with Fura-3/AM. Results: There was an inverse correlation between Cx43 expression and MC proliferation (P<0.05). SLDT studies revealed that there was no difference in the gap junction channel function between the normal and Aldosterone (Aldo)-stimulated groups (P>0.05). Our data also showed that the mineralcorticoid receptor (MR) antagonist spironolactone, ERK1/2 inhibitor PD98059 and PKC inhibitor GF109203X could attenuate the down-regulation of Cx43 expression in Aldo-induced MC proliferation; however, the PI3K inhibitor LY294002 could block MC proliferation without affecting Cx43 expression at either the mRNA or protein level. In addition, Aldo promoted MC proliferation in parallel with increasing [Ca²⁺]_i (P<0.05), suggesting that the classical PKC pathway might be activated. Conclusions: Our study provides preliminary evidence that Cx43 is an important regulator of Aldo-promoted MC proliferation. Furthermore, reduced Cx43 expression promoted MC proliferation independent of the gap junction channel function, and this process might be mediated through the ERK1/2- and PKC-dependent pathways.

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Coculture of Embryonic Ventricular Myocytes and Mouse Embryonic Stem Cell Enhance Intercellular Signaling by Upregulation of Connexin43

Cited by 6 publications

References 33 publications

Exploiting AT2R to Improve CD117 Stem Cell Function In Vitro and In Vivo - Perspectives for Cardiac Stem Cell Therapy

Exploiting AT2R to Improve CD117 Stem Cell Function In Vitro and In Vivo - Perspectives for Cardiac Stem Cell Therapy

Co-culture with neonatal cardiomyocytes enhances the proliferation of iPSC-derived cardiomyocytes via FAK/JNK signaling

Beyond Gap Junction Channel Function: the Expression of Cx43 Contributes to Aldosterone-Induced Mesangial Cell Proliferation via the ERK1/2 and PKC Pathways

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