Differentiation of Rat Bone Marrow Mesenchymal Stem Cells into Adipocytes and Cardiomyocytes after Treatment with Platelet Lysate
Abstract
Background: Mesenchymal stem cells (MSCs) are multipotential cells and their therapeutic potency is under intense investigation. Studying the effect of different induction factors on MSCs could increase our knowledge about the differentiation potency of these cells. One of the most important sources of these factors in mammalian body is platelet. Platelet lysate (PL) contains many growth factors and therefore it can be used as a differentiation inducer. In the present study, the effect of PL on differentiation of rat bone marrow MSCs into cardiomyocytes was studied.
Materials Methods: To study the differentiation-inducing effect of PL, MSCs were treated with 2.5, 5 and 10% PL. Early results of this study showed that PL in high concentrations (10%) induces adipogenic differentiation of MSCs. Therefore, to evaluate differentiation to cardiomyocytes, MSCs were cultured in media containing lower levels of PL (2.5% and 5%) and then cardiomyogenic differentiation was induced by treatment with 5-azacytidine. Differentiation of MSCs was evaluated using direct observation of beating cells, immunostaining and real-time PCR techniques.
Results: The results of qPCR showed that treatment with PL alone increased the expression of cardiac alpha actinin (CAA) being predictable by earlier observation of beating cells in PL-treated groups. The results of staining assays against cardiac alpha actinin also showed that there were stained cells in PL-treated groups.
Conclusion: The results of the present study showed that PL is a powerful induction factor for differentiation of MSCs into different cell lines such as cardiomyocytes and adipocytes.
Dimmeler S, Burchfield J, Zeiher AM. Cell-based therapy of myocardial infarction. Arterioscler Thromb Vasc Biol. 2008; 28:208-16.
Boyle AJ, Schulman SP, Hare JM, et al. Is stem cell therapy ready for patients? Stem Cell Therapy for Cardiac Repair. Ready for the Next Step. Circulation. 2006; 114: 339-52.
Kadam S, Govindasamy V, Bhonde R. Generation of functional islets from human umbilical cord and placenta derived mesenchymal stem cells. Methods Mol Biol. 2012; 879:291-313.
Jia HY, Yu JH, Lei TC. The multi-lineage differentiation and angiogenesis ability of adipose-derived adult mesenchymal stem cells. Clin Lab. 2014; 60: 1747-55.
Hashimoto R, Katoh Y, Miyamoto Y, et al. Increased extracellular and intracellular Ca(2+) lead to adipocyte accumulation in bone marrow stromal cells by different mechanisms. Biochem Biophys Res Commun. 2015 Feb 20; 457(4): 647-52.
Murphy MK, Huey DJ, Hu JC, et al. TGF-β1, GDF-5, and BMP-2 stimulation induces chondrogenesis in expanded human articular chondrocytes and marrow-derived stromal cells. Stem Cells. 2014 Mar; 33(3):762-73.
Zheng YH, Xiong W, Su K, et al. Multilineage differentiation of human bone marrow mesenchymal stem cells in vitro and in vivo. Exp Ther Med. 2013; 5:1576-80.
Cui X, Chen L, Xue T, et al. Human umbilical cord and dental pulp-derived mesenchymal stem cells: Biological characteristics and potential roles in vitro and in vivo. Mol Med Rep. 2015 May; 11(5):3269-78.
Struys T, Moreels M, Martens W, et al. Ultrastructural and immunocytochemical analysis of multilineage differentiated human dental pulp- and umbilical cord-derived mesenchymal stem cells. Cells Tissues Organs. 2011; 193:366-78.
Fukuhara S, Tomita S, Yamashiro S, et al. Direct cell-cell interaction of cardiomyocytes is key for bone marrow stromal cells to go into cardiac lineage in vitro. J Thorac Cardiovasc Surg. 2003; 125: 1470-80.
Behfar A, Yamada S, Crespo-Diaz R, et al. Guided cardiopoiesis enhances therapeutic benefit of bone marrow human mesenchymal stem cells in chronic myocardial infarction. J Am Coll Cardiol. 2010; 56:721-34.
Sachinidis A, Gissel C, Nierhoff D, et al. Identification of plateled-derived growth factor-BB as cardiogenesis-inducing factor in mouse embryonic stem cells under serum-free conditions. Cell Physiol Biochem. 2003; 13: 423-9.
Xaymardan M, Tang L, Zagreda L, et al. Platelet-derived growth factor-AB promotes the generation of adult bone marrow-derived cardiac myocytes. Circulation Research. 2004; 94: E39-45.
Hupkes M, Jonsson MK, Scheenen WJ, et al. Epigenetics: DNA demethylation promotes skeletal myotube maturation. FASEB J. 2011; 25: 3861-72.
Rosca AM, Burlacu A. Effect of 5-azacytidine: evidence for alteration of the multipotent ability of mesenchymal stem cells. Stem Cells Dev. 2011; 20: 1213-21.
van den Dolder J, Mooren R, Vloon AP, et al. Platelet-rich plasma: quantification of growth factor levels and the effect on growth and differentiation of rat bone marrow cells. Tissue Eng. 2006; 12: 3067-73.
Kilian O, Flesch I, Wenisch S, et al. Effects of platelet growth factors on human mesenchymal stem cells and human endothelial cells in vitro. Eur J Med Res. 2004; 9: 337-44.
Fekete N, Gadelorge M, Furst D, et al. Platelet lysate from whole blood-derived pooled platelet concentrates and apheresis-derived platelet concentrates for the isolation and expansion of human bone marrow mesenchymal stromal cells: production process, content and identification of active components. Cytotherapy. 2012; 14: 540-54.
Shim WS, Jiang S, Wong P, et al. Ex vivo differentiation of human adult bone marrow stem cells into cardiomyocyte-like cells. Biochem Biophys Res Commun. 2004; 324: 481-8.
Bittira B, Kuang JQ, Al-Khaldi A, et al. In vitro preprogramming of marrow stromal cells for myocardial regeneration. Ann Thorac Surg. 2002; 74: 1154-9; discussion 9-60.
Kardami E. Stimulation and inhibition of cardiac myocyte proliferation in vitro. Mol Cell Biochem. 1990; 92: 129-35.
Choi WY, Gemberling M, Wang J, et al. In vivo monitoring of cardiomyocyte proliferation to identify chemical modifiers of heart regeneration. Development. 2013; 140:660-6.
Vantler M, Karikkineth BC, Naito H, et al. PDGF-BB protects cardiomyocytes from apoptosis and improves contractile function of engineered heart tissue. J Mol Cell Cardiol. 2010; 48:1316-23.
Tang QQ, Lane MD. Adipogenesis: from stem cell to adipocyte. Annu Rev Biochem. 2012; 81:715-36.
Rangappa S, Fen C, Lee EH, et al. Transformation of adult mesenchymal stem cells isolated from the fatty tissue into cardiomyocytes. Ann Thorac Surg. 2003; 75: 775-9.
Fukuda K. Development of regenerative cardiomyocytes from mesenchymal stem cells for cardiovascular tissue engineering. Artif Organs. 2001; 25: 187-93.
Liu Y, Song J, Liu W, et al. Growth and differentiation of rat bone marrow stromal cells: does 5-azacytidine trigger their cardiomyogenic differentiation? Cardiovasc Res. 2003; 58: 460-8.
Ishimine H, Yamakawa N, Sasao M, et al. N-Cadherin is a prospective cell surface marker of human mesenchymal stem cells that have high ability for cardiomyocyte differentiation. Biochem Biophys Res Commun. 2013; 438: 753-9.
Li H, Zuo S, Pasha Z, et al. GATA-4 promotes myocardial transdifferentiation of mesenchymal stromal cells via up-regulating IGFBP-4. Cytotherapy. 2011; 13:1057-65.
Koninckx R, Hensen K, Daniëls A, et al. Human bone marrow stem cells co-cultured with neonatal rat cardiomyocytes display limited cardiomyogenic plasticity. Cytotherapy. 2009; 11: 778-92.
Lachaud CC, López-Beas J, Soria B, et al. EGF-induced adipose tissue mesothelial cells undergo functional vascular smooth muscle differentiation. Cell Death Dis. 2014; 5: e1304.
Shi ZD, Abraham G, Tarbell JM. Shear stress modulation of smooth muscle cell marker genes in 2-D and 3-D depends on mechanotransduction by heparan sulfate proteoglycans and ERK1/2. PLoS One. 2010; 5: e12196.
Singh K, Cassano M, Planet E, et al. A KAP1 phosphorylation switch controls MyoD function during skeletal muscle differentiation. Genes Dev. 2015; 29: 513-25.
Choi JH, Song YJ, Lee H. The histone demethylase KDM4B interacts with MyoD to regulate myogenic differentiation in C2C12 myoblast cells. Biochem Biophys Res Commun. 2015; 456:872-8.
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Issue | Vol 10, No 1 (2016) | |
Section | Articles | |
Keywords | ||
Platelet lysate Cardiomyogenic differentiation 5-azacytidine Mesenchymal stem cells |
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