Autonomous control of expression of genes for insulin-like growth factors during the proliferation and differentiation of C2C12 mouse myoblasts in serum-free culture

Yoshiko, Yuji; Hirao, Keita; Sakabe, Kou; Seiki, Kanji; Takezawa, Junichi; Maeda, Norihiko

doi:10.1016/s0024-3205(96)00547-4

Cited by 17 publications

(12 citation statements)

References 31 publications

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“…Several reports exist on serum-free culture media designed for the analysis of myogenesis using several species of animal cells such as chick embryo (22)(23)(24), rat primary satellite cells (25), rat myoblast cell line (26,27), mouse myoblast cell line (28,29), and human primary satellite cells (30 -32). Those reports focused mainly on cell proliferation, DNA synthesis, morphological differentiation, or creatine phosphokinase activity, but not on secretory proteins during myogenesis.…”

Section: Discussionmentioning

confidence: 99%

“…In all reports, inoculation of the cells was performed in the presence of serum and some of the culture media contained biological fluids or unknown components. C2C12 myoblast cells, which were established by Yaffe and used as one of the common model system for the analysis of myogenic differentiation, were cultured and induced to differentiate under serum-free conditions (28). However, in their culture system, both serum supplement present at cell inoculation and adaptation period for serum-free environments were required, and unknown components were contained in the medium.…”

Section: Discussionmentioning

confidence: 99%

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Serum-Free Culture Conditions for Analysis of Secretory Proteinases during Myogenic Differentiation of Mouse C2C12 Myoblasts

Goto

Miyazaki

Funabiki

et al. 1999

Analytical Biochemistry

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Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Serum-Free Culture Conditions for Analysis of Secretory Proteinases during Myogenic Differentiation of Mouse C2C12 Myoblasts

Goto

Miyazaki

Funabiki

et al. 1999

Analytical Biochemistry

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“…For some time, researchers have tested serum-free media for the study of myoblasts in culture [Florini and Roberts, 1979;Dollenmeier et al, 1981;Yoshiko et al, 1996;Minotto et al, 1998;Goto et al, 1999]. In each of these studies, a medium was found that effectively supports proliferation and differentiation in a single myoblast cell line.…”

Section: Introductionmentioning

confidence: 99%

Differentiation of Myoblasts in Serum-Free Media: Effects of Modified Media Are Cell Line-Specific

Lawson¹,

Purslow²

2000

Cells Tissues Organs

137

101

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Myoblast cell lines are grown and differentiated readily in cell culture. Two cell lines typically used for investigating the growth and differentiation of muscle are the mouse cell line C2C12 and the rat cell line L6. The differentiation of these cells in vitro requires a switch from a serum-rich medium to a less rich medium after the cells have reached confluence. Since the components present in serum are not well characterized, the use of a better defined medium for these studies was investigated. C2C12 and L6 myoblasts were differentiated in both serum-containing and serum-free media. The differentiation state of these cultures was then tested both microscopically and biochemically. Cultures were checked for myotube formation, the activity of creatine phosphokinase and the presence of sarcomeric actin. In C2C12 cells, the extent of differentiation was greater in the serum-free than in the serum-containing system. In both media types, the C2C12 cells produced sarcomeric actin, showing the presence of sarcomere structure in the myotubes. In L6 cells, however, myotubes were readily formed in medium containing 2% horse serum, but not in the serum-free system. In addition, the ability of C2C12 cells to differentiate on substrates coated with extracellular matrix proteins was shown to be media-dependent. The presence of extracellular matrix proteins did not enable L6 cells to form myotubes when cultured in serum-free media. Primary cultures of chick myoblasts were able to differentiate in both media tested, with Dulbecco’s modified Eagle medium containing horse serum being a more efficient medium for cell fusion. This study shows a divergence in muscle cell line responses in three cell lines, two of which are typically used as ‘model systems’ for understanding muscle growth and development.

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“…It is known that various growth factors contained in serum play a role in the proliferation of cells. Cells continuously proliferate under high serum concentrations (10%), while low serum concentrations (1%) permit their terminal differentiation (30,31,38). Under low serum culturing of SPTL cells, the induction of thyroid differentiation marker genes other than Nkx2-1 was not observed, although the expression of Foxe1, Nis (Scl5a5), and Tpo mRNAs appeared to have slightly increased toward the end of six days in culture.…”

Section: Discussionmentioning

confidence: 99%

“…1D and see below). When the serum concentration was changed to 2%, which switches cells from proliferation to differentiation (30,31), the expression of mRNA encoding SCA1 decreased, while mRNAs encoding NKX2-1 increased during 6 days of 2% FBS culture (Fig. 3A).…”

Section: Generation Of a Mouse Thyroid Sp Cell-derived Cell Linementioning

confidence: 99%

An Adult Mouse Thyroid Side Population Cell Line that Exhibits Enriched Epithelial–Mesenchymal Transition

Murata

Iwadate

Takizawa

et al. 2017

Thyroid

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Background: Studies of thyroid stem/progenitor cells have been hampered due to the small organ size and lack of tissue, which limits the yield of these cells. A continuous source that allows the study and characterization of thyroid stem/progenitor cells is desired to push the field forward. Method: A cell line was established from Hoechst-resistant side population cells derived from mouse thyroid that were previously shown to contain stem/progenitor-like cells. Characterization of these cells were carried out by using in vitro two-and three-dimensional cultures and in vivo reconstitution of mice after orthotopic or intravenous injection, in conjunction with quantitative reverse transcription polymerase chain reaction, Western blotting, immunohisto(cyto)chemistry/immunofluorescence, and RNA seq analysis. Results: These cells were named SPTL (side population cell-derived thyroid cell line). Under low serum culturing conditions, SPTL cells expressed the thyroid differentiation marker NKX2-1, a transcription factor critical for thyroid differentiation and function, while no expression of other thyroid differentiation marker genes were observed. SPTL cells formed follicle-like structures in Matrigel Ò cultures, which did not express thyroid differentiation marker genes. In mouse models of orthotopic and intravenous injection, the latter following partial thyroidectomy, a few SPTL cells were found in part of the follicles, most of which expressed NKX2-1. SPTL cells highly express genes involved in epithelial-mesenchymal transition, as demonstrated by RNA seq analysis, and exhibit a gene-expression pattern similar to anaplastic thyroid carcinoma.Conclusion: These results demonstrate that SPTL cells have the capacity to differentiate into thyroid to a limited degree. SPTL cells may provide an excellent tool to study stem cells, including cancer stem cells of the thyroid.

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Autonomous control of expression of genes for insulin-like growth factors during the proliferation and differentiation of C2C12 mouse myoblasts in serum-free culture

Cited by 17 publications

References 31 publications

Serum-Free Culture Conditions for Analysis of Secretory Proteinases during Myogenic Differentiation of Mouse C2C12 Myoblasts

Serum-Free Culture Conditions for Analysis of Secretory Proteinases during Myogenic Differentiation of Mouse C2C12 Myoblasts

Differentiation of Myoblasts in Serum-Free Media: Effects of Modified Media Are Cell Line-Specific

An Adult Mouse Thyroid Side Population Cell Line that Exhibits Enriched Epithelial–Mesenchymal Transition

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