Involvement of racl in calcium-dependent activation of JNK and cell injury by oxidative stress in the rat myoblast cell line, H9c2

Geigeria poisoning in sheep, locally known as ‘vermeersiekte’, is an economically important plant poisoning in southern Africa. The toxic principles contained by the toxic plants are believed to be several sesquiterpene lactones, such as geigerin, vermeeric acid and vermeerin, which cause striated muscle lesions in small stock. Because of ethical issues surrounding the use of live animals in toxicity studies, there is currently a dire need to establish an in vitro model that can be used to replace traditional animal experimentation. The objective of this study was to determine the cytotoxicity of geigerin in a murine myoblast cell line (C2C12) using methyl-thiazol-tetrazolium (MTT) and lactate dehydrogenase (LDH) assays, annexin V and propidium iodide (PI) flow cytometry and transmission electron microscopy (TEM). Mouse myoblasts were exposed to 2.0 mM, 2.5 mM and 5.0 mM geigerin for 24, 48 and 72 h. A concentration-dependent cytotoxic response was observed. Apoptosis was detected by means of annexin V flow cytometry during the first 24 h and apoptotic bodies were also visible on TEM. According to the LDH and PI flow cytometry results, myoblast cell membranes were not injured. We concluded that the murine myoblast cell line (C2C12) is a suitable model for future studies planned to evaluate the cytotoxicity of other and combinations of sesquiterpene lactones, with and without metabolic activation, implicated in ‘vermeersiekte’ and to elucidate the subcellular effects of these myotoxins on cultured myoblasts.

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Glucose Regulated Production of Human Insulin in Genetically Modified Myoblast Cell Line (C2C12)

Journal of Korean Endocrine Society ◽

10.3803/jkes.2006.21.6.526 ◽

2006 ◽

Vol 21 (6) ◽

pp. 526

Author(s):

Seungjoon Oh ◽

Jeong-Taek Woo ◽

Sung Woon Kim ◽

Jin-Woo Kim ◽

Young Seol Kim

Keyword(s):

Cell Line ◽

Human Insulin ◽

Genetically Modified ◽

Myoblast Cell Line ◽

Myoblast Cell

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Propofol elicits autophagy via endoplasmic reticulum stress and calcium exchange in C2C12 myoblast cell line

PLoS ONE ◽

10.1371/journal.pone.0197934 ◽

2018 ◽

Vol 13 (5) ◽

pp. e0197934 ◽

Cited By ~ 4

Author(s):

Xi Chen ◽

Long-Yun Li ◽

Jin-Lan Jiang ◽

Kai Li ◽

Zhen-Bo Su ◽

...

Keyword(s):

Endoplasmic Reticulum ◽

Endoplasmic Reticulum Stress ◽

Cell Line ◽

C2c12 Myoblast ◽

Myoblast Cell Line ◽

Calcium Exchange ◽

Myoblast Cell

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Isolation and characterization of a variant myoblast cell line that is temperature sensitive for differentiation

Molecular and Cellular Biology ◽

10.1128/mcb.8.6.2335-2341.1988 ◽

1988 ◽

Vol 8 (6) ◽

pp. 2335-2341

Author(s):

R J Akhurst ◽

N B Flavin ◽

J Worden

Keyword(s):

Cell Line ◽

Progenitor Cell ◽

Temperature Shift ◽

Temperature Sensitive ◽

Isolation And Characterization ◽

New Variant ◽

Myoblast Cell Line ◽

Myoblast Cell ◽

Progenitor Cell Line

A new variant rat myogenic cell line, ts485, was isolated by subcloning the cell line ts3b2 (H. T. Nguyen, R. M. Medford, and B. Nadal-Ginard, Cell 34:281-293, 1983). Unlike the progenitor cell line, ts485 was thermosensitive for differentiation. Experiments with conditioned medium suggested that diffusible extracellular factors were not involved in dictating the differential phenotypes of ts485 cells cultured at the permissive and nonpermissive temperatures. Temperature shift experiments performed on cultures of ts485 cells indicated that the temperature-sensitive lesion was in a factor active during the growth phase and required to trigger a cascade of events leading to terminal differentiation.

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A Theacrine-Based Supplement Increases Cellular NAD+ Levels and Affects Biomarkers Related to Sirtuin Activity in C2C12 Muscle Cells In Vitro

Nutrients ◽

10.3390/nu12123727 ◽

2020 ◽

Vol 12 (12) ◽

pp. 3727

Author(s):

Petey W. Mumford ◽

Shelby C. Osburn ◽

Carlton D. Fox ◽

Joshua S. Godwin ◽

Michael D. Roberts

Keyword(s):

Cell Line ◽

Mitochondrial Biogenesis ◽

C2c12 Myoblast ◽

Mrna Levels ◽

Nicotinamide Phosphoribosyltransferase ◽

Protein Levels ◽

Myoblast Cell Line ◽

Rodent Cell ◽

Myoblast Cell

There is evidence in rodents to suggest that theacrine-based supplements modulate tissue sirtuin activity as well as other biological processes associated with aging. Herein, we examined if a theacrine-based supplement (termed NAD3) altered sirtuin activity in vitro while also affecting markers of mitochondrial biogenesis. The murine C2C12 myoblast cell line was used for experimentation. Following 7 days of differentiation, myotubes were treated with 0.45 mg/mL of NAD3 (containing ~2 mM theacrine) for 3 and 24 h (n = 6 treatment wells per time point). Relative to control (CTL)-treated cells, NAD3 treatments increased (p < 0.05) Sirt1 mRNA levels at 3 h, as well as global sirtuin activity at 3 and 24 h. Follow-up experiments comparing 24 h NAD3 or CTL treatments indicated that NAD3 increased nicotinamide phosphoribosyltransferase (NAMPT) and SIRT1 protein levels (p < 0.05). Cellular nicotinamide adenine dinucleotide (NAD+) levels were also elevated nearly two-fold after 24 h of NAD3 versus CTL treatments (p < 0.001). Markers of mitochondrial biogenesis were minimally affected. Although these data are limited to select biomarkers in vitro, these preliminary findings suggest that a theacrine-based supplement can modulate select biomarkers related to NAD+ biogenesis and sirtuin activity. However, these changes did not drive increases in mitochondrial biogenesis. While promising, these data are limited to a rodent cell line and human muscle biopsy studies are needed to validate and elucidate the significance of these findings.

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