Cell Culture Models and Methods

Nikolskaya, Alena; Sharma, Vinod

doi:10.1007/978-1-4419-6658-2_10

Cited by 5 publications

(5 citation statements)

References 47 publications

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“…Indeed, Bloch et al recently demonstrated that the concentration of MED12 mutant cells gradually decreases in the primary culture of MED12-LMs even without passaging (26). The study by Bloch et al also suggested that the loss of MED12 mutant cells is partially due to the detachment of mutant cells from the culture flask, which likely reflects the weaker adherence of SMCs to the plastic surface compared to fibroblasts (40). Then, the subsequent passaging further dilutes SMCs in MED12-LM cell culture (25).…”

Section: Discussionmentioning

confidence: 99%

Subtype-Specific Tumor-Associated Fibroblasts Contribute to the Pathogenesis of Uterine Leiomyoma

Wu¹,

Serna²,

Thomas³

et al. 2017

Cancer Research

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Recent genomic studies have identified subtypes of uterine leiomyoma (LM) with distinctive genetic alterations. Here we report the elucidation of the biological characteristics of the two most prevalent LM subtypes, MED12 mutant (MED12-LM) and HMGA2-overexpressing (HMGA2-LM) LM. Since each tumor carries only one genetic alteration, both subtypes are considered to be monoclonal. Approximately 90% of cells in HMGA2-LM were smooth muscle cells (SMC) with HMGA2 overexpression. In contrast, MED12-LM consisted of similar numbers of SMC and non-SMC, which were mostly tumor-associated fibroblasts (TAF). Paradoxically, TAF carried no mutations in MED12, suggesting an interaction between SMC and TAF to coordinate their growth. The higher amount of ECM in MED12-LM than HMGA2-LM was partially due to the high concentration of collagen-producing TAF. SMC growth in a xenograft assay was driven by progesterone in both LM subtypes. In contrast, TAF in MED12-LM proliferated in response to estradiol, whereas progesterone had no effect. The high concentration of estrogen-responsive TAF in MED12-LM explains the inconsistent discoveries between in vivo and in vitro studies on the mitogenic effect of estrogen and raises questions regarding the accuracy of previous studies utilizing MED12-LM cell culture. In addition, the differential effects of estradiol and progesterone on these LM subtypes emphasize the importance of subtypes and genotypes in designing non-surgical therapeutic strategies for LM.

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Section: Discussionmentioning

confidence: 99%

Subtype-Specific Tumor-Associated Fibroblasts Contribute to the Pathogenesis of Uterine Leiomyoma

Wu¹,

Serna²,

Thomas³

et al. 2017

Cancer Research

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“…The cardiomyoblast H9C2 cell line was purchased from ATCC (CRL‐1446). Neonatal rat ventricular myocytes (NRVMs) were primarily isolated from neonatal (1–2 day‐old) Sprague–Dawley rat hearts .…”

Section: Methodsmentioning

confidence: 99%

The alteration of protein prenylation induces cardiomyocyte hypertrophy through Rheb–mTORC1 signalling and leads to chronic heart failure

Guan

Chen

et al. 2015

The Journal of Pathology

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G protein-regulated cell function is crucial for cardiomyocytes, and any deregulation of its gene expression or protein modification can lead to pathological cardiac hypertrophy. Herein, we report that protein prenylation, a lipidic modification of G proteins that facilitates their association with the cell membrane, might control the process of cardiomyocyte hypertrophy. We found that geranylgeranyl diphosphate synthase (GGPPS), a key enzyme involved in protein prenylation, played a critical role in postnatal heart growth by regulating cardiomyocyte size. Cardiac-specific knockout of GGPPS in mice led to spontaneous cardiac hypertrophy, beginning from week 4, accompanied by the persistent enlargement of cardiomyocytes. This hypertrophic effect occurred by altered prenylation of G proteins. Evaluation of the prenylation, membrane association and hydrophobicity showed that Rheb was hyperactivated and increased mTORC1 signalling pathway after GGPPS deletion. Protein farnesylation or mTORC1 inhibition blocked GGPPS knockdown-induced mTORC1 activation and suppressed the larger neonatal rat ventricle myocyte size and cardiomyocyte hypertrophy in vivo, demonstrating a central role of the FPP-Rheb-mTORC1 axis for GGPPS deficiency-induced cardiomyocyte hypertrophy. The sustained cardiomyocyte hypertrophy progressively provoked cardiac decompensation and dysfunction, ultimately causing heart failure and adult death. Importantly, GGPPS was down-regulated in the hypertrophic hearts of mice subjected to transverse aortic constriction (TAC) and in failing human hearts. Moreover, HPLC-MS/MS detection revealed that the myocardial farnesyl diphosphate (FPP):geranylgeranyl diphosphate (GGPP) ratio was enhanced after pressure overload. Our observations conclude that the alteration of protein prenylation promotes cardiomyocyte hypertrophic growth, which acts as a potential cause for pathogenesis of heart failure and may provide a new molecular target for hypertrophic heart disease clinical therapy.

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“…Their diameters are in the 7–9 μm range [54,69,70], and their membrane resistances are an order of magnitude higher than in fibroblasts assessed in situ [52,55]. …”

Section: What Is Different In a Dishmentioning

confidence: 99%

Fibroblast–myocyte coupling in the heart: Potential relevance for therapeutic interventions

Ongstad

Kohl

2016

Journal of Molecular and Cellular Cardiology

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Cardiac myocyte-fibroblast electrotonic coupling is a well-established fact in vitro. Indirect evidence of its presence in vivo exists, but few functional studies have been published. This review describes the current knowledge of fibroblast-myocyte electrical signalling in the heart. Further research is needed to understand the frequency and extent of heterocellular interactions in vivo in order to gain a better understanding of their relevance in healthy and diseased myocardium. It is hoped that associated insight into myocyte-fibroblast coupling in the heart may lead to the discovery of novel therapeutic targets and the development of agents for improving outcomes of myocardial scarring and fibrosis. This article is part of a special issue entitled “Exploring Fibrosis as the Next Target for Myocardial Remodeling.”

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Cell Culture Models and Methods

Cited by 5 publications

References 47 publications

Subtype-Specific Tumor-Associated Fibroblasts Contribute to the Pathogenesis of Uterine Leiomyoma

Subtype-Specific Tumor-Associated Fibroblasts Contribute to the Pathogenesis of Uterine Leiomyoma

The alteration of protein prenylation induces cardiomyocyte hypertrophy through Rheb–mTORC1 signalling and leads to chronic heart failure

Fibroblast–myocyte coupling in the heart: Potential relevance for therapeutic interventions

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