Development of a clonal equine myoblast cell line capable of terminal differentiation into mature myotubes in vitro

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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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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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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Antisense Rb RNA Inhibits in Vitro Myogenic Differentiation in a Myoblast Cell Line, C2

Animal Cell Technology: Basic & Applied Aspects ◽

10.1007/978-94-011-5161-0_8 ◽

1998 ◽

pp. 41-44

Author(s):

Masayuki Kobayashi ◽

Yukika Yamauchi ◽

Akiko Tanaka

Keyword(s):

Cell Line ◽

Myogenic Differentiation ◽

Myoblast Cell Line ◽

Myoblast Cell

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Iso-Mukaadial Acetate from Warburgia salutaris Enhances Glucose Uptake in the L6 Rat Myoblast Cell Line

Biomolecules ◽

10.3390/biom9100520 ◽

2019 ◽

Vol 9 (10) ◽

pp. 520 ◽

Cited By ~ 1

Author(s):

Nontokozo Z. Msomi ◽

Francis O. Shode ◽

Ofentse J. Pooe ◽

Sithandiwe Mazibuko-Mbeje ◽

Mthokozisi B. C. Simelane

Keyword(s):

Protein Kinase ◽

Glucose Uptake ◽

Cell Line ◽

In Vitro Assays ◽

Dependent Manner ◽

Scavenging Activity ◽

Glucose Utilisation ◽

Myoblast Cell Line ◽

Myoblast Cell

Diabetes mellitus (DM) is a chronic metabolic disorder which has become a major risk to the health of humankind, as its global prevalence is increasing rapidly. Currently available treatment options in modern medicine have several adverse effects. Thus, there is an urgent need to develop alternative cost-effective, safe, and active treatments for diabetes. In this regard, medicinal plants provide the best option for new therapeutic remedies desired to be effective and safe. Recently, we focused our attention on drimane sesquiterpenes as potential sources of antimalarial and antidiabetic agents. In this study, iso-mukaadial acetate (Iso) (1), a drimane-type sesquiterpenoid from the ground stem bark of Warburgia salutaris, was investigated for glucose uptake enhancement in the L6 rat myoblast cell line. In vitro assays with L6 skeletal muscle cells were used to test for cytotoxicity, glucose utilisation, and western blot analysis. Additionally, the inhibition of carbohydrate digestive enzymes and 1,1-diphenyl-2- picrylhydrazyl (DPPH) scavenging activity were analysed in vitro. The cell viability effect of iso-mukaadial acetate was the highest at 3 µg/mL with a percentage of 98.4. Iso-mukaadial acetate also significantly and dose-dependently increased glucose utilisation up to 215.18% (12.5 µg/mL). The increase in glucose utilisation was accompanied by enhanced 5’ adenosine monophosphate-activated protein kinase (AMPK)and protein kinase B (AKT) in dose-dependent manner. Furthermore, iso-mukaadial acetate dose-dependently inhibited the enzymes α-amylase and α-glucosidase. Scavenging activity against DPPH was displayed by iso-mukaadial acetate in a concentration-dependent manner. The findings indicate the apparent therapeutic efficacy of iso-mukaadial acetate isolated from W. salutaris as a potential new antidiabetic agent.

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Cell cycle withdrawal without concomitant differentiation: Analysis of a G1-specific temperature-sensitive murine myoblast cell line

Developmental Biology ◽

10.1016/0012-1606(88)90394-6 ◽

1988 ◽

Vol 129 (2) ◽

pp. 476-494 ◽

Cited By ~ 7

Author(s):

Reid S. Compton ◽

Irwin R. Konigsberg

Keyword(s):

Cell Cycle ◽

Cell Line ◽

Temperature Sensitive ◽

Myoblast Cell Line ◽

Specific Temperature ◽

Myoblast Cell

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Lentivector-Mediated Transfer of Bmi-1 and Telomerase in Muscle Satellite Cells Yields a Duchenne Myoblast Cell Line with Long-Term Genotypic and Phenotypic Stability

Human Gene Therapy ◽

10.1089/104303403322495034 ◽

2003 ◽

Vol 14 (16) ◽

pp. 1525-1533 ◽

Cited By ~ 66

Author(s):

Christophe Cudré-Mauroux ◽

Teresa Occhiodoro ◽

Stéphane König ◽

Patrick Salmon ◽

Laurent Bernheim ◽

...

Keyword(s):

Cell Line ◽

Satellite Cells ◽

Phenotypic Stability ◽

Muscle Satellite Cells ◽

Myoblast Cell Line ◽

Myoblast Cell

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Structure and expression of ferritin genes in a human promyelocytic cell line that differentiates in vitro

Molecular and Cellular Biology ◽

10.1128/mcb.6.2.566-573.1986 ◽

1986 ◽

Vol 6 (2) ◽

pp. 566-573

Author(s):

C C Chou ◽

R A Gatti ◽

M L Fuller ◽

P Concannon ◽

A Wong ◽

...

Keyword(s):

Cell Line ◽

Terminal Differentiation ◽

Amino Acid Sequences ◽

Multigene Families ◽

Cdna Clones ◽

Macrophage Differentiation ◽

Time Points ◽

Human Ferritin ◽

Complex Manner

HL-60 is a human promyelocytic cell line with the capability of differentiating in vitro to give neutrophils, macrophages, or eosinophils. We screened libraries of HL-60 cDNA clones representing different time points during these differentiation processes to isolate clones corresponding to mRNAs whose expression is regulated during terminal differentiation. Upon sequencing this group of regulated clones, one clone encoding the heavy subunit and two clones encoding the light subunit of human ferritin were identified by reference to published amino acid sequences. Southern blot analyses showed that these clones are encoded by distinct multigene families. These clones identify two mRNAs whose ratios vary in a complex manner during both neutrophil and macrophage differentiation.

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