Establishment of myoblast cell line and identification of key genes regulating myoblast differentiation in a marine teleost, Sebastes schlegelii

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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Cartap-induced cytotoxicity in mouse C2C12 myoblast cell line and the roles of calcium ion and oxidative stress on the toxic effects

Toxicology ◽

10.1016/j.tox.2005.11.002 ◽

2006 ◽

Vol 219 (1-3) ◽

pp. 73-84 ◽

Cited By ~ 22

Author(s):

Jiunn-Wang Liao ◽

Jaw-Jou Kang ◽

Chian-Ren Jeng ◽

Shao-Kuang Chang ◽

Ming-Jang Kuo ◽

...

Keyword(s):

Oxidative Stress ◽

Cell Line ◽

Calcium Ion ◽

Toxic Effects ◽

C2c12 Myoblast ◽

Myoblast Cell Line ◽

Myoblast Cell ◽

And Oxidative Stress

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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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Creation and characterization of an immortalized canine myoblast cell line: Myok9

Mammalian Genome ◽

10.1007/s00335-020-09833-5 ◽

2020 ◽

Vol 31 (3-4) ◽

pp. 95-109

Author(s):

Sara Mata López ◽

Cynthia Balog-Alvarez ◽

Emily H. Canessa ◽

Yetrib Hathout ◽

Kristy J. Brown ◽

...

Keyword(s):

Cell Line ◽

Myoblast Cell Line ◽

Myoblast Cell

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Cytoprotective potential of anti-ischemic drugs against chemotherapy-induced cardiotoxicity in H9c2 myoblast cell line

Acta Pharmaceutica ◽

10.2478/acph-2013-0038 ◽

2013 ◽

Vol 63 (4) ◽

pp. 493-503 ◽

Cited By ~ 4

Author(s):

Tiam Feridooni ◽

Chris Mac Donald ◽

Di Shao ◽

Pollen Yeung ◽

Remigius U. Agu

Keyword(s):

Cell Death ◽

Cell Line ◽

Cancer Drug ◽

H9c2 Cell ◽

Drug Induced ◽

Cardiac Model ◽

Myoblast Cell Line ◽

Myoblast Cell ◽

H9c2 Cell Line

Abstract To investigate potential prevention or attenuation of anti- cancer drug induced cardiotoxicity using anti-ischemic drugs, a rat myoblast (H9c2) cell line was used as our in vitro cardiac model. Irinotecan and doxorubicin were found to be cytotoxic for the H9c2 cell line with IC50 of 30.69 ± 6.20 and 20.94 ± 6.05 mmol L-1, respectively. 5-Flurouracil and cladribine were not cytotoxic and thus IC50 could not be calculated. When 100 mmol L-1 doxorubicin was incubated for 72 hours with 50 mmol L-1 diltiazem, 100 mmol L-1 dexrazoxane and 100 mmol L-1 losartan, respectively, there was a 58.7 ± 10.2, 52.2 ± 11.7 and 44.7 ± 5.4 % reduction in cell death. When 200 mmol L-1 irinotecan was incubated for 72 hours with 100 mmol L-1 dexrazoxane, losartan and diltiazem, respectively, a 27.7 ± 6.9, 25.6 ± 5.1, and 19.1 ± 2.3 % reduction in cell death was observed. Our data suggests that losartan and diltiazem were as effective as dexrazoxane in protecting the cells against irinotecan- and doxorubicin-induced cell toxicity. These findings offer potential uses of anti- -ischemic drugs for ablation of cytotoxicity in response to mitochondrial injury, thereby improving patient outcomes and reducing health-care costs.

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