The Quality of Porcine Mesenchymal Stem Cells and Their Osteo- and Adipogenic Cell Derivatives – The Level of Proapoptotic Bad Protein Expression / Jakość Mezenchymalnych Komórek Macierzystych Świni Oraz Ich Pochodnych Zróżnicowanych W Kierunku Komórek Szeregu Osteo- I Adipogennego – Poziom Ekspresji Proapoptotycznego Białka Bad

Open access

Abstract

The aim of the research was to evaluate the quality of porcine mesenchymal stem cells (MSCs) and MSC-derived osteoblasts/osteocytes (bone cells) and adipocytes (fat cells). This evaluation was performed on the basis of molecular analysis for proapoptotic BAD protein expression. MSCs isolated from the pig bone marrow were cultured in vitro for five weeks in three types of culture media: differentiating towards the osteoblasts/osteocytes (O) and adipocytes (A) and non-differentiating, control medium (C). In all groups of cells, the relative extent of BAD protein expression was estimated by western blotting. Significant differences in the posttranslational abundance of BAD proteins were noted between MSCs differentiated into the osteogenic and the adipogenic cell lineages (P<0.05). Summarizing the results, we conclude that posttranslational level of BAD protein expression can be used as a reliable marker for assessing the quality of both MSCs and their cell derivatives. Interestingly, the semi-quantitative profile of BAD protein expression in differentiated cells turned out to be lower than that observed in undifferentiated cells, demonstrating that the culture conditions used for pro-osteogenic or pro-adipogenic cellular transformation did not affect negatively the quality of MSCs.

If the inline PDF is not rendering correctly, you can download the PDF file here.

  • Alfaro M.P. Vincent A. Saraswati S. Thorne C.A. Hong C.C. Lee E. Young P.P. (2010). s FRP2 suppression of bone morphogenic protein (BMP) and Wnt signaling mediates mesenchymal stem cell (MSC) self-renewal promoting engraftment and myocardial repair. J. Biol. Chem. 285: 35645-35653.

  • Antonsson B. Conto F. Ciavatta A.M. Montesuit S. Lewus S. Martinou I. Bernasconi L. Bernard A. Mermod J.C. Mazzei G. Maundrell K. Gambal e F. Sadoul R. Martinou J.C. (1997). Inhibition of Bax channel forming activity by Bcl-2. Science 277: 330-372.

  • Ashkenazi A. Dixit V.M. (1998). Death receptors: signaling and modulation. Science 281: 1305-1308.

  • Bajek A. Olkowska J. Drewa T. (2011). Mesenchymal stem cells asatherapeutic tool in tissue and organ regeneration (in Polish). Adv. Hyg. Exp. Med. 65: 124-132.

  • Bieback K. Wuchter P. Besser D. Franke W. Becker M. Ott M. Pacher M. Ma N. Stamm C. Klüter H. Müller A. Ho A.D. START-MSCconsortium (2012). Mesenchymal stromal cells (MSCs): science and f(r)iction. J. Mol. Med. (Berl). 90: 773-782.

  • Bosch P. Pratt S.L. Stice S.L. (2006). Isolation characterization gene modification and nuclear reprogramming of porcine mesenchymal stem cells. Biol. Reprod. 74: 46-57.

  • Cai A. Zheng D. Dong Y. Qiu R. Huang Y. Song Y. Jiang Z. Rao S. Liao X. Kuang J. Dai G. Mai W. (2011). Efficacy of Atorvastatin combined with adipose-derived mesenchymal stem cell transplantation on cardiac function in rats with acute myocardial infarction. Acta. Biochim. Biophys. 43: 857-866.

  • Caplan A.I. (1991). Mesenchymal stem cells. J. Orthop. Res. 9: 641-650.

  • Chipuk J.E. Green D.R. (2005). Do inducers of apoptosis trigger caspase-independent cell death? Nat. Rev. Mol. Cell Biol. 6: 268-275.

  • Coleman C.M. Vaughan E.E. Browe D.C. Mooney E. Howard L. Barry F.P. (2013). Growth differentiation factor-5 enhances in vitro mesenchymal stromal cell chondrogenesis and hypertrophy. Stem Cells Dev. doi:10.1089/scd.2012.0282.

  • Datta S.R. Dudek H. Tao X. Masters S. Fu H. Gotoh Y. Greenberg M.E. (1997). Akt phosphorylation of BADcouples survival signals to the cell-intrinsic death machinery. Cell 91: 231-241.

  • Dominici M. Le Blanc K. Mueller I. Slaper - Cortenbach I. Marini F. Krause D. Deans R. Keating A. Prockop Dj. Horwitz E. (2006). Minimal criteria for defining multipotent mesenchymal stromal cells. The International Society for Cellular Therapy position statement. Cytotherapy 8: 315-317.

  • Hanada M. Feng J. Hemmings B.A. (2004). Structure regulation and function of PKB/AKT - a major therapeutic target. Biochim. Biophys. Acta 1697: 3-16.

  • Huppeertz B. Frank H.G. Kaufmann P. (1999). The apoptosis cascade morphological and immunohistochemical methods for its visualization. Anat. Embryol. 200: 1-18.

  • Izadpanah R. Kaushal D. Kriedt C. Tsien F. Patel B. Dufour J. Bunnell B.A. (2008). Long-term in vitro expansion alters the biology of adult mesenchymal stem cells. Cancer Res. 68: 4229-4238.

  • Juhásová J. Juhás Š. Klíma J. Strnádel J. Holubová M. Motlík J. (2011). Osteogenic differentiation of miniature pig mesenchymal stem cells in 2Dand 3Denvironment. Physiol. Res. 60: 559-571.

  • Kidd V.J. Lahti J.M. Teitz T. (2000). Proteolytic regulation of apoptosis. Seminars Cell Develop. Biol. 11: 191-201.

  • Krześlak A. (2010). Akt kinase:akey regulator of metabolism and progression of tumors. Adv. Hyg. Exp. Med. (in Polish). 64: 490-503.

  • Laemmli U.K. (1970). Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227: 680-685.

  • Mao J.J. Marion N.W. (2009). Essential Stem Cell Methods. Editors Lanza R. Klimanskaya I. Academic Press 1st ed. 297 pp.

  • Opiela J. Kątska- Książkiewicz L. Lipiński D. Słomski R. Bzowska M. Ryńska B. (2008 a). Interactions among activity of glucose-6-phosphate dehydrogenase in immature oocytes expression of apoptosis-related genes Bcl-2 and Bax and developmental competence following IVPin cattle. Theriogenology 69: 546-555.

  • Opiela J. Kątska - Książkiewicz L. Smorąg Z. (2008 b). DNAanalysis: theory and practice (in Polish). Słomski R. (ed.). University of Life Sciences Poznań pp. 530-536.

  • Quevedo H.C. Hatzistergos K.E. Oskouei B.N. Feigenbaum G.S. Rodri- guez J.E. Valdes D. Pattany P.M. Zambrano J.P. Hu Q. Mc Niece I. Held - man A.W. Hare J.M. (2009). Allogeneic mesenchymal stem cells restore cardiac function in chronic ischemic cardiomyopathy via trilineage differentiating capacity. Proc. Natl. Acad. Sci. 106: 14022-14027.

  • Ratajczak M.Z. Kucia M. (2005). Stem cells - the challenge of the 21st century? (in Polish). Postępy Biol. Kom. 23: 11-26.

  • Ringe J. Kaps C. Schmitt B. Büscher K. Bartel J. Smolian H. Schultz O. Bur- mester G.R. Häupl T. Sittinger M. (2002). Porcine mesenchymal stem cells. Induction of distinct mesenchymal cell lineages. Cell Tissue Res. 307: 321-327.

  • Słupianek A. Pytel D. Majsterek I. (2007). The role of oncogenic tyrosine kinases in the cellular response to anticancer therapy (in Polish). Adv. Hyg. Exp. Med. 61: 819-827.

  • Stępień A. Izdebska M. Grzanka A. (2007). Types of cell death (in Polish). Adv. Hyg. Exp. Med. 61: 420-428.

  • Szalata M. Pławski A. Słomski R. (2008). DNAanalysis: theory and practice (in Polish). Słomski R. (ed.). University of Life Sciences Poznań pp. 503-511.

  • Urbaniak - Kujda D. Wołowiec D. Tomaszewska- Toporska B. Kapelko- - Słowik K. Kuliczkowski K. (2005). Mesenchymal stem cells: their biology and prospects for clinical applications. Acta Haematol. Pol. 2: 161-166.

  • Vaux D.L. Cory S. Adams J.M. (1988). Bcl-2 gene promotes haemopoietic cell survival and cooperates with c-myc to immortalize pre-Bcells. Nature 335: 440-442.

Search
Journal information
Impact Factor

IMPACT FACTOR 2018: 1.515
5-year IMPACT FACTOR: 1.246

CiteScore 2018: 1.4

SCImago Journal Rank (SJR) 2018: 0.509
Source Normalized Impact per Paper (SNIP) 2018: 0.869

Metrics
All Time Past Year Past 30 Days
Abstract Views 0 0 0
Full Text Views 209 90 3
PDF Downloads 89 54 1