Abstract
Cell- and tissue culture methodology form an important base for biological, biochemical and biomedical research. Most cell culture techniques require the use of animal sera for the successful propagation of cells. However, the varying composition between batches has fuelled the need for alternatives. In the case of serum derived from animal foetuses, ethical concerns have also been raised. Here we compare the use of a platelet derived lysate (Plysate), which is currently under development as a serum substitute, in the culturing of primary human muscle cells to foetal bovine serum (FBS). In cells cultured with Plysate, differentiation into myotubes, glucose-uptake, phosphatidylinositol 3-kinase (PI3K) activity and expression and phosphorylation of ERK1/2 MAPK and PKB/Akt was impaired. Thus for primary human skeletal muscle Plysate is a sub-optimal substitute for FBS.
Keywords: Foetal bovine serum (FBS), Insulin action, Platelet derived lysate (Plysate), Primary human skeletal muscle cell culture, Serum replacement
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References
- Al-Khalili L., Chibalin A.V., Kannisto K., Zhang B.B., Permert J., Holman G.D., Ehrenborg E., Ding V.D., Zierath J.R., Krook A. Insulin action in cultured human skeletal muscle cells during differentiation: assessment of cell surface GLUT4 and GLUT1 content. Cell Mol. Life Sci. 2003;60:991–998. doi: 10.1007/s00018-003-3001-3. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Al-Khalili L., Krämer D., Wretenberg P., Krook A. Human skeletal muscle cell differentiation is associated with changes in myogenic markers and enhanced insulin-mediated MAPK and PKB phosphorylation. Acta Physiol. Scand. 2004;180:395–403. doi: 10.1111/j.1365-201X.2004.01259.x. [DOI] [PubMed] [Google Scholar]
- Cha B.S., Ciaraldi T.P., Park K.S., Carter L., Mudaliar S.R. and Henry R.R. 2005. Impaired fatty acid metabolism in type 2 diabetic skeletal muscle is reversed by PPAR{gamma} agonists. Am. J. Physiol. Endocrinol. Metab. [DOI] [PubMed]
- Donna S., Mamchaoui K., Cooper R.N., Seigneurin-Venin S., Tremblay J., Butler-Browne G.S., Mouly V. Telomerase can extend the proliferative capacity of human myoblasts, but does not lead to their immortalization. Mol. Cancer Res. 2003;1:643–653. [PubMed] [Google Scholar]
- Fiaschi T., Chiarugi P., Buricchi F., Giannoni E., Taddei M.L., Magnelli L., Cozzi G., Raugei G., Ramponi G. Down-regulation of platelet-derived growth factor receptor signaling during myogenesis. Cell Mol. Life Sci. 2003;60:2721–2735. doi: 10.1007/s00018-003-3293-3. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Fukamizu H., Grinnell F. Spatial organization of extracellular matrix and fibroblast activity: effects of serumtransforming growth factor betaand fibronectin. Exp. Cell Res. 1990;190:276–282. doi: 10.1016/0014-4827(90)90197-I. [DOI] [PubMed] [Google Scholar]
- Gstraunthaler G. Alternatives to the use of fetal bovine serum: serum-free cell culture. Altex. 2003;20:275–281. [PubMed] [Google Scholar]
- Jayme D.W., Blackman K.E. Culture media for propagation of mammalian cells, viruses, and other biologicals. Adv Biotechnol Processes. 1985;5:1–30. [PubMed] [Google Scholar]
- Jochems C.E., Valk J.B., Stafleu F.R., Baumans V. The use of fetal bovine serum: ethical or scientific problem? Altern. Lab Anim. 2002;30:219–227. doi: 10.1177/026119290203000208. [DOI] [PubMed] [Google Scholar]
- Krook A., Wallberg-Henriksson H., Zierath J.R. Sending the Signal: Molecular Mechanisms Regulating Glucose Uptake. Med. Sci. Sports Exerc. 2004;36:1212–1217. doi: 10.1249/01.MSS.0000132387.25853.3B. [DOI] [PubMed] [Google Scholar]
- Kwon M.S., Dojima T. and Park E.Y. 2004. Use of plant-derived protein hydrolysates for enhancing growth of Bombyx mori insect cells in suspension culture. Biotechnol. Appl. Biochem. [DOI] [PubMed]
- Lebkowski J.S., Schain L.R., Okarma T.B. Serum-free culture of hematopoietic stem cells: a review. Stem Cells. 1995;13:607–612. doi: 10.1002/stem.5530130605. [DOI] [PubMed] [Google Scholar]
- Liselott Johansson J.K., Olov H. and Ohlson S. 2003. Platelet lysate: a replacement for fetal bovine serum in animal cell culture? Cytotechnology 67–74. [DOI] [PMC free article] [PubMed]
- Mumford J.H., Carnes D.L., Cochran D.L., Oates T.W. The effects of platelet-derived growth factor-BB on periodontal cells in an in vitro wound model. J. Periodontol. 2001;72:331–340. doi: 10.1902/jop.2001.72.3.331. [DOI] [PubMed] [Google Scholar]
- Olov H. and Westermark B. 1989. Blood Platelet Lysatemethod of its preparation and a cell culture medium containing blood platelet lysate. European Patent Cooperation 413727.
- Olov H. and Westermark B. 2003. Preparation of a blood platelet lysate for use in a cell culture medium for hybridoma cells. Patent USA.
- Peng H., Huard J. Muscle-derived stem cells for musculoskeletal tissue regeneration and repair. Transpl. Immunol. 2004;12:311–319. doi: 10.1016/j.trim.2003.12.009. [DOI] [PubMed] [Google Scholar]
- Perez-Reyes N., Halbert C.L., Smith P.P., Benditt E.P., McDougall J.K. Immortalization of primary human smooth muscle cells. PNAS. 1992;89:1224–1228. doi: 10.1073/pnas.89.4.1224. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Ross R., Bowen-Pope D.F., Raines E.W. Platelet-derived growth factor and its role in health and disease. Philos. Trans. R. Soc. Lond. B Biol. Sci. 1990;327:155–169. doi: 10.1098/rstb.1990.0051. [DOI] [PubMed] [Google Scholar]
- Sporn M.B., Roberts A.B. Transforming growth factor-beta. Multiple actions and potential clinical applications. JAMA. 1989;262:938–941. doi: 10.1001/jama.262.7.938. [DOI] [PubMed] [Google Scholar]
- Tallquist M., Kazlauskas A. PDGF signaling in cells and mice. Cytokine Growth Factor Rev. 2004;15:205–213. doi: 10.1016/j.cytogfr.2004.03.003. [DOI] [PubMed] [Google Scholar]
- Valk J., Mellor D., Brands R., Fischer R., Gruber F., Gstraunthaler G., Hellebrekers L., Hyllner J., Jonker F.H., Prieto P., Thalen M., Baumans V. The humane collection of fetal bovine serum and possibilities for serum-free cell and tissue culture. Toxicol. In Vitro. 2004;18:1–12. doi: 10.1016/j.tiv.2003.08.009. [DOI] [PubMed] [Google Scholar]
