C2C12

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C2C12 myotubes under light microscope, 10x magnification C2C12 cells x10 mag.jpg
C2C12 myotubes under light microscope, 10x magnification

C2C12 is an immortalized mouse myoblast cell line. The C2C12 cell line is a subclone of myoblasts that were originally obtained by Yaffe and Saxel at the Weizmann Institute of Science in Israel in 1977. [1] Developed for in vitro studies of myoblasts isolated from the complex interactions of in vivo conditions, C2C12 cells are useful in biomedical research. [2] These cells are capable of rapid proliferation under high serum conditions and differentiation into myotubes under low serum conditions. Mononucleated myoblasts can later fuse to form multinucleated myotubes under low serum conditions or starvation, leading to the precursors of contractile skeletal muscle cells in the process of myogenesis. [3] C2C12 cells are used to study the differentiation of myoblasts, osteoblasts, and myogenesis, to express various target proteins, and to explore mechanistic biochemical pathways.

Contents

Morphology

Wild-type C2C12 cells have a radial branching morphology consisting of long fibers extending in many directions. C2C12 cells can be cultured in a variety of conditions to induce specific responses of interest. For example, assisted by the cell line's high differentiation rate and fusion rate, fibronectin templates can be micro-plated to petri dishes or cell culture flasks in order to induce specific growth patterns, such as that of skeletal muscle cell interactions with extracellular matrix components. [4] The introduction of adhesion molecules can alter the growth pattern of C2C12 cells to a longitudinal distribution exhibiting polarity. [5] There are many ways to regulate the shape of C2C12 myoblasts genetically and environmentally, from stress, to cytoskeleton alteration, to growth factors. The scaffolding of C2C12 cells is particularly important for studying muscle tissue regeneration post-injury or after tissue wasting due to disease or ICU rehabilitation.

Uses in research

C2C12 cells have been shown to effectively incorporate exogenous cDNA and nucleic acids by transfection. In the piloting research originally conducted by Yaffe and Saxel, C2C12 were obtained through serial passage of myoblasts cultured from the thigh muscle of C3H mice after crush injury. In their study, a set of C2C12 cells were cultured from normal mouse myoblasts, which were cultured from two-month-old C3H mice after crush injury. Within two days, the normal cells differentiated into spindle-shaped mononucleated myoblasts. After four days, multinucleated myotube networks formed, and a few days after, sarcomeres and Z-lines could be observed. [6] In contrast, the dystrophic cells formed shortened fibers covered in fibroblasts, a hallmark of muscle wasting. [1]

C2C12 cells demonstrate rapid development and maturation into functional skeletal muscle cells or cardiac muscle cells, having the ability to contract and generate force. [6] The rate of muscle formation from C2C12 cells can be controlled by the introduction of loss-of-functions genes vital for the fusion of myoblasts and myogenesis. [7] Under necrotic conditions, such as tumor necrosis factor alpha (TNF-α), direct protein loss, particularly myosin heavy chain protein, in C2C12 skeletal muscle cells has been shown. [8] C2C12 cells were used to elucidate inactivated X chromosome (Xi) replication during early S-phase of the cell cycle and is regulated epigenetically. [9] C2C12 cells are especially convenient for studying the cell cycle due to its high division rate.

Related Research Articles

A syncytium or symplasm is a multinucleate cell that can result from multiple cell fusions of uninuclear cells, in contrast to a coenocyte, which can result from multiple nuclear divisions without accompanying cytokinesis. The muscle cell that makes up animal skeletal muscle is a classic example of a syncytium cell. The term may also refer to cells interconnected by specialized membranes with gap junctions, as seen in the heart muscle cells and certain smooth muscle cells, which are synchronized electrically in an action potential.

<span class="mw-page-title-main">Muscle cell</span> Type of cell found in muscle tissue

A muscle cell, also known as a myocyte, is a mature contractile cell in the muscle of an animal. In humans and other vertebrates there are three types: skeletal, smooth, and cardiac (cardiomyocytes). A skeletal muscle cell is long and threadlike with many nuclei and is called a muscle fiber. Muscle cells develop from embryonic precursor cells called myoblasts.

<span class="mw-page-title-main">MyoD</span> Mammalian protein found in Homo sapiens

MyoD, also known as myoblast determination protein 1, is a protein in animals that plays a major role in regulating muscle differentiation. MyoD, which was discovered in the laboratory of Harold M. Weintraub, belongs to a family of proteins known as myogenic regulatory factors (MRFs). These bHLH transcription factors act sequentially in myogenic differentiation. Vertebrate MRF family members include MyoD1, Myf5, myogenin, and MRF4 (Myf6). In non-vertebrate animals, a single MyoD protein is typically found.

Myosatellite cells, also known as satellite cells, muscle stem cells or MuSCs, are small multipotent cells with very little cytoplasm found in mature muscle. Satellite cells are precursors to skeletal muscle cells, able to give rise to satellite cells or differentiated skeletal muscle cells. They have the potential to provide additional myonuclei to their parent muscle fiber, or return to a quiescent state. More specifically, upon activation, satellite cells can re-enter the cell cycle to proliferate and differentiate into myoblasts.

<span class="mw-page-title-main">Myogenesis</span> Formation of muscular tissue, particularly during embryonic development

Myogenesis is the formation of skeletal muscular tissue, particularly during embryonic development.

<span class="mw-page-title-main">Myogenin</span> Mammalian protein found in Homo sapiens

Myogenin, is a transcriptional activator encoded by the MYOG gene. Myogenin is a muscle-specific basic-helix-loop-helix (bHLH) transcription factor involved in the coordination of skeletal muscle development or myogenesis and repair. Myogenin is a member of the MyoD family of transcription factors, which also includes MyoD, Myf5, and MRF4.

<span class="mw-page-title-main">Mesoangioblast</span>

A mesoangioblast is a type of progenitor cell that is associated with vasculature walls. Mesoangioblasts exhibit many similarities to pericytes, which are found in the small vessels. Mesoangioblasts are multipotent stem cells with the potential to progress down the endothelial or mesodermal lineages. Mesoangioblasts express the critical marker of angiopoietic progenitors, KDR (FLK1). Because of these properties, mesoangioblasts are a precursor of skeletal, smooth, and cardiac muscle cells along with endothelial cells. Research has suggested their application for stem cell therapies for muscular dystrophy and cardiovascular disease.

<span class="mw-page-title-main">Muscle</span> Basic biological tissue present in animals

Muscle is a soft tissue, one of the four basic types of animal tissue. Muscle tissue gives skeletal muscles the ability to contract. Muscle is formed during embryonic development, in a process known as myogenesis. Muscle tissue contains special contractile proteins called actin and myosin which interact to cause movement. Among many other muscle proteins present are two regulatory proteins, troponin and tropomyosin.

<span class="mw-page-title-main">Actin, alpha skeletal muscle</span> Protein-coding gene in the species Homo sapiens

Actin, alpha skeletal muscle is a protein that in humans is encoded by the ACTA1 gene.

<span class="mw-page-title-main">ADAM12</span> Human protein-coding gene

Disintegrin and metalloproteinase domain-containing protein 12 is an enzyme that in humans is encoded by the ADAM12 gene. ADAM12 has two splice variants: ADAM12-L, the long form, has a transmembrane region and ADAM12-S, a shorter variant, is soluble and lacks the transmembrane and cytoplasmic domains.

<span class="mw-page-title-main">Integrin alpha 7</span>

Alpha-7 integrin is a protein that in humans is encoded by the ITGA7 gene. Alpha-7 integrin is critical for modulating cell-matrix interactions. Alpha-7 integrin is highly expressed in cardiac muscle, skeletal muscle and smooth muscle cells, and localizes to Z-disc and costamere structures. Mutations in ITGA7 have been associated with congenital myopathies and noncompaction cardiomyopathy, and altered expression levels of alpha-7 integrin have been identified in various forms of muscular dystrophy.

<span class="mw-page-title-main">TNFRSF12A</span> Protein-coding gene in the species Homo sapiens

Tumor necrosis factor receptor superfamily member 12A also known as the TWEAK receptor (TWEAKR) is a protein that in humans is encoded by the TNFRSF12A gene.

<span class="mw-page-title-main">CDH15</span> Protein-coding gene in the species Homo sapiens

Cadherin-15 is a protein that in humans is encoded by the CDH15 gene.

<span class="mw-page-title-main">P19 cell</span>

P19 cells is an embryonic carcinoma cell line derived from an embryo-derived teratocarcinoma in mice. The cell line is pluripotent and can differentiate into cell types of all three germ layers. Also, it is the most characterized embryonic carcinoma (EC) cell line that can be induced into cardiac muscle cells and neuronal cells by different specific treatments. Indeed, exposing aggregated P19 cells to dimethyl sulfoxide (DMSO) induces differentiation into cardiac and skeletal muscle. Also, exposing P19 cells to retinoic acid (RA) can differentiate them into neuronal cells.

A myokine is one of several hundred cytokines or other small proteins and proteoglycan peptides that are produced and released by skeletal muscle cells in response to muscular contractions. They have autocrine, paracrine and/or endocrine effects; their systemic effects occur at picomolar concentrations.

<span class="mw-page-title-main">MYF5</span> Protein-coding gene in the species Homo sapiens

Myogenic factor 5 is a protein that in humans is encoded by the MYF5 gene. It is a protein with a key role in regulating muscle differentiation or myogenesis, specifically the development of skeletal muscle. Myf5 belongs to a family of proteins known as myogenic regulatory factors (MRFs). These basic helix loop helix transcription factors act sequentially in myogenic differentiation. MRF family members include Myf5, MyoD (Myf3), myogenin, and MRF4 (Myf6). This transcription factor is the earliest of all MRFs to be expressed in the embryo, where it is only markedly expressed for a few days. It functions during that time to commit myogenic precursor cells to become skeletal muscle. In fact, its expression in proliferating myoblasts has led to its classification as a determination factor. Furthermore, Myf5 is a master regulator of muscle development, possessing the ability to induce a muscle phenotype upon its forced expression in fibroblastic cells.

<span class="mw-page-title-main">MYF6</span> Protein-coding gene in the species Homo sapiens

Myogenic factor 6 is a protein that in humans is encoded by the MYF6 gene. This gene is also known in the biomedical literature as MRF4 and herculin. MYF6 is a myogenic regulatory factor (MRF) involved in the process known as myogenesis.

<span class="mw-page-title-main">Calponin 3, acidic</span> Protein-coding gene in the species Homo sapiens

Calponin 3. acidic is a protein that in humans is encoded by the CNN3 gene.

<span class="mw-page-title-main">MiR-206</span>

MiR-206 is a microRNA with a sequence conserved across most mammalian species, and in humans is a member of the myo-miR family of miRNAs, which includes miR-1, miR-133, and miR-208a/b. Mir-206 is well established for the regulation of cellular processes involving skeletal muscle development, as well as mitochondrial functioning. miR-206 is studied in C2C12 myoblast cells as this is a widely used model for the study of cellular differentiation of skeletal muscle. The biogenesis of miR-206 is unique in that the primary mature transcript is generated from the 3p arm of the precursor microRNA hairpin rather than the 5p arm. Currently, miR-206 has approximately twelve miRNA family members, and the cognate seed sequence of the miR-206 family is conserved across all twelve miRNA members.

Entomoculture is the subfield of cellular agriculture which specifically deals with the production of insect tissue in vitro. It draws on principles more generally used in tissue engineering and has scientific similarities to Baculovirus Expression Vectors or soft robotics. The field has mainly been proposed because of its potential technical advantages over mammalian cells in generating cultivated meat. The name of the field was coined by Natalie Rubio at Tufts University.

References

  1. 1 2 Yaffe, David; Saxel, Ora (22 December 1977). "Serial passaging and differentiation of myogenic cells isolated from dystrophic mouse muscle". Nature. 270 (5639): 725–727. Bibcode:1977Natur.270..725Y. doi:10.1038/270725a0. ISSN   0028-0836. PMID   563524. S2CID   4196110.
  2. ↑ "C2C12 Cell Line" . Retrieved 12 July 2018.
  3. ↑ "Working with the C2C12 cell line". Research in Myogenesis. 4 February 2012. Retrieved 3 May 2017.
  4. ↑ Bajaj, Piyush; Reddy, Bobby; Millet, Larry; Wei, Chunan; Zorlutuna, Pinar; Bao, Gang; Bashir, Rashid (1 September 2011). "Patterning the differentiation of C2C12 skeletal myoblasts". Integrative Biology. 3 (9): 897–909. doi:10.1039/c1ib00058f. ISSN   1757-9708. PMID   21842084.
  5. ↑ Mermelstein, C. S. (5 May 2003). "Changes in cell shape, cytoskeletal proteins and adhesion sites of cultured cells after extracellular Ca2+ chelation" (PDF). Brazilian Journal of Medical and Biological Research. 36 (8): 1111–1116. doi: 10.1590/s0100-879x2003000800018 . PMID   12886466.
  6. 1 2 McMahon, D. K.; Anderson, P. A.; Nassar, R.; Bunting, J. B.; Saba, Z.; Oakeley, A. E.; Malouf, N. N. (1 June 1994). "C2C12 cells: biophysical, biochemical, and immunocytochemical properties". American Journal of Physiology. Cell Physiology. 266 (6): C1795–C1802. doi:10.1152/ajpcell.1994.266.6.c1795. ISSN   0363-6143. PMID   8023908.
  7. ↑ Bi, Pengpeng; Ramirez-Martinez, Andres; Li, Hui; Cannavino, Jessica; McAnally, John R.; Shelton, John M.; Sánchez-Ortiz, Efrain; Bassel-Duby, Rhonda; Olson, Eric N. (21 April 2017). "Control of muscle formation by the fusogenic micropeptide myomixer". Science. 356 (6335): 323–327. Bibcode:2017Sci...356..323B. doi:10.1126/science.aam9361. ISSN   1095-9203. PMC   5502127 . PMID   28386024.
  8. ↑ Li, Y. P.; Schwartz, R. J.; Waddell, I. D.; Holloway, B. R.; Reid, M. B. (1 July 1998). "Skeletal muscle myocytes undergo protein loss and reactive oxygen-mediated NF-kappaB activation in response to tumor necrosis factor alpha". FASEB Journal. 12 (10): 871–880. doi: 10.1096/fasebj.12.10.871 . ISSN   0892-6638. PMID   9657527.
  9. ↑ Casas-Delucchi, Corella S.; Brero, Alessandro; Rahn, Hans-Peter; Solovei, Irina; Wutz, Anton; Cremer, Thomas; Leonhardt, Heinrich; Cardoso, M. Cristina (1 March 2011). "Histone acetylation controls the inactive X chromosome replication dynamics". Nature Communications. 2: 222. Bibcode:2011NatCo...2..222C. doi:10.1038/ncomms1218. ISSN   2041-1723. PMC   3072080 . PMID   21364561.
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