DOI QR코드

DOI QR Code

Alteration of Apoptosis during Differentiation in Human Dental Pulp-Derived Mesenchymal Stem Cell

  • Lee, Hyeon-Jeong (College of Veterinary Medicine, Gyeongsang National University) ;
  • Park, Byung-Joon (College of Veterinary Medicine, Kyungpook National University) ;
  • Jeon, Ryoung-Hoon (College of Veterinary Medicine, Gyeongsang National University) ;
  • Jang, Si-Jung (College of Veterinary Medicine, Gyeongsang National University) ;
  • Son, Young-Bum (College of Veterinary Medicine, Gyeongsang National University) ;
  • Lee, Sung-Lim (College of Veterinary Medicine, Gyeongsang National University) ;
  • Rho, Gyu-Jin (College of Veterinary Medicine, Gyeongsang National University) ;
  • Kim, Seung-Joon (College of Veterinary Medicine, Kyungpook National University) ;
  • Lee, Won-Jae (College of Veterinary Medicine, Kyungpook National University)
  • Received : 2019.03.18
  • Accepted : 2019.03.26
  • Published : 2019.03.31

Abstract

Because mesenchymal stem cells (MSCs) maintain distinct capacities with respect to self-renewal, differentiation ability and immunomodulatory function, they have been highly considered as the therapeutic agents for cell-based clinical application. Of particular, differentiation condition alters characteristics of MSCs, including cellular morphology, expression of gene/protein and cell surface molecule, immunological property and apoptosis. However, the previous results for differentiation-related apoptosis in MSCs have still remained controversial due to varied outcomes. Therefore, the present study aimed to disclose periodical alterations of pro- and anti-apoptosis in MSCs under differentiation inductions. The human dental pulp-derived MSCs (DP-MSCs) were differentiated into adipocytes and osteoblasts during early (1 week), middle (2 weeks) and late (3 weeks) stages, and were investigated on their apoptosis-related changes by Annexin V assay, qRT-PCR and western blotting. The ratio of apoptotic cell population was significantly (p < 0.05) elevated during the early to middle stages of differentiations but recovered up to the similar level of undifferentiated state at the late stage of differentiation. In the expression of mRNA and protein, whereas expressions of pro-apoptosis-related makers (BAX and BAK) were not altered in any kind and duration of differentiation inductions, anti-apoptosis marker (BCL2) was significantly (p < 0.05) elevated even at the early stage of differentiations. The recovery of apoptotic cell population at the late stage of differentiation is expected to be associated with the response by elevation of anti-apoptotic molecules. The present study may contribute on understanding for cellular mechanism in differentiation of MSCs and provide background data in clinical application of MSCs in the animal biotechnology to develop effective and safe therapeutic strategy.

Keywords

References

  1. Blagosklonny MV. 2003. Apoptosis, proliferation, differentiation: in search of the order. Semin. Cancer Biol. 13:97-105. https://doi.org/10.1016/S1044-579X(02)00127-X
  2. Constance CM, Morgan JI 4th, Umek RM. 1996. C/EBPalpha regulation of the growth-arrest-associated gene gadd45. Mol. Cell Biol. 16:3878-3883. https://doi.org/10.1128/MCB.16.7.3878
  3. Giansanti V, Torriglia A, Scovassi AI. 2011. Conversation between apoptosis and autophagy: "Is it your turn or mine?". Apoptosis 16:321-333. https://doi.org/10.1007/s10495-011-0589-x
  4. Graneli C, Thorfve A, Ruetschi U, Brisby H, Thomsen P, Lindahl A, Karlsson C. 2014. Novel markers of osteogenic and adipogenic differentiation of human bone marrow stromal cells identified using a quantitative proteomics approach. Stem Cell Res. 12:153-165. https://doi.org/10.1016/j.scr.2013.09.009
  5. Gross A, McDonnell JM, Korsmeyer SJ. 1999. BCL-2 family members and the mitochondria in apoptosis. Genes Dev. 13:1899-1911. https://doi.org/10.1101/gad.13.15.1899
  6. Inoue N, Yahagi N, Yamamoto T, Ishikawa M, Watanabe K, Matsuzaka T, Nakagawa Y, Takeuchi Y, Kobayashi K, Takahashi A, Suzuki H, Hasty AH, Toyoshima H, Yamada N, Shimano H. 2008. Cyclin-dependent kinase inhibitor, p21WAF1/CIP1, is involved in adipocyte differentiation and hypertrophy, linking to obesity, and insulin resistance. J. Biol. Chem. 283:21220-21229. https://doi.org/10.1074/jbc.M801824200
  7. Kumar BM, Maeng GH, Lee YM, Kim TH, Lee JH, Jeon BG, Ock SA, Yoo JG, Rho GJ. 2012. Neurogenic and cardiomyogenic differentiation of mesenchymal stem cells isolated from minipig bone marrow. Res. Vet. Sci. 93:749-757. https://doi.org/10.1016/j.rvsc.2011.09.012
  8. Lee WJ, Hah YS, Ock SA, Lee JH, Jeon RH, Park JS, Lee SI, Rho NY, Rho GJ, Lee SL. 2015. Cell source-dependent in vivo immunosuppressive properties of mesenchymal stem cells derived from the bone marrow and synovial fluid of minipigs. Exp. Cell Res. 333:273-288. https://doi.org/10.1016/j.yexcr.2015.03.015
  9. Liu F, Akiyama Y, Tai S, Maruyama K, Kawaguchi Y, Muramatsu K, Yamaguchi K. 2008. Changes in the expression of CD106, osteogenic genes, and transcription factors involved in the osteogenic differentiation of human bone marrow mesenchymal stem cells. J. Bone Miner. Metab. 26:312-320. https://doi.org/10.1007/s00774-007-0842-0
  10. Lo Furno D, Graziano AC, Caggia S, Perrotta RE, Tarico MS, Giuffrida R, Cardile V. 2013. Decrease of apoptosis markers during adipogenic differentiation of mesenchymal stem cells from human adipose tissue. Apoptosis 18:578-588. https://doi.org/10.1007/s10495-013-0830-x
  11. Marzo I, Brenner C, Zamzami N, Jurgensmeier JM, Susin SA, Vieira HL, Prevost MC, Xie Z, Matsuyama S, Reed JC, Kroemer G. 1998. Bax and adenine nucleotide translocator cooperate in the mitochondrial control of apoptosis. Science 281:2027-2031. https://doi.org/10.1126/science.281.5385.2027
  12. Ock SA, Baregundi SR, Lee YM, Lee JH, Jeon RH, Lee SL, Park JK, Hwang SC, Rho GJ. 2016a. Comparison of Immunomodulation Properties of Porcine Mesenchymal Stromal/Stem Cells Derived from the Bone Marrow, Adipose Tissue, and Dermal Skin Tissue. Stem Cells Int. 2016:9581350. https://doi.org/10.1155/2016/9581350
  13. Ock SA, Lee YM, Park JS, Shivakumar SB, Moon SW, Sung NJ, Lee WJ, Jang SJ, Park JM, Lee SC, Lee SL, Rho GJ. 2016b. Evaluation of phenotypic, functional and molecular characteristics of porcine mesenchymal stromal/stem cells depending on donor age, gender and tissue source. J. Vet. Med. Sci. 78:987-995. https://doi.org/10.1292/jvms.15-0596
  14. Oliver L, Hue E, Rossignol J, Bougras G, Hulin P, Naveilhan P, Heymann D, Lescaudron L, Vallette FM. 2011. Distinct roles of Bcl-2 and Bcl-Xl in the apoptosis of human bone marrow mesenchymal stem cells during differentiation. PLoS One 6:e19820. https://doi.org/10.1371/journal.pone.0019820
  15. Oltvai ZN, Milliman CL, Korsmeyer SJ. 1993. Bcl-2 heterodimerizes in vivo with a conserved homolog, Bax, that accelerates programmed cell death. Cell 74:609-619. https://doi.org/10.1016/0092-8674(93)90509-O
  16. Orelio C, Dzierzkak E. 2007. Bcl-2 expression and apoptosis in the regulation of hematopoietic stem cells. Leuk. Lymph. 48:16-24. https://doi.org/10.1080/10428190601032529
  17. Parsch D, Fellenberg J, Brummendorf TH, Eschlbeck AM, Richter W. 2004. Telomere length and telomerase activity during expansion and differentiation of human mesenchymal stem cells and chondrocytes. J. Mol. Med. (Berl) 82:49-55. https://doi.org/10.1007/s00109-003-0506-z
  18. Ragni E, Vigano M, Rebulla P, Giordano R, Lazzari L. 2013. What is beyond a qRT-PCR study on mesenchymal stem cell differentiation properties: how to choose the most reliable housekeeping genes. J. Cell Mol. Med. 17:168-180. https://doi.org/10.1111/j.1582-4934.2012.01660.x
  19. Sun HJ, Bahk YY, Choi YR, Shim JH, Han SH, Lee JW. 2006. A proteomic analysis during serial subculture and osteogenic differentiation of human mesenchymal stem cell. J. Orthop. Res. 24:2059-2071. https://doi.org/10.1002/jor.20273
  20. Ullah I, Choe YH, Khan M, Bharti D, Shivakumar SB, Lee HJ, Son YB, Shin Y, Lee SL, Park BW, Ock SA, Rho GJ. 2018. Dental pulp-derived stem cells can counterbalance peripheral nerve injury-induced oxidative stress and supraspinal neuro-inflammation in rat brain. Sci. Rep. 8:15795. https://doi.org/10.1038/s41598-018-34151-x
  21. Wang CY, Chen LL, Kuo PY, Chang JL, Wang YJ, Hung SC. 2010. Apoptosis in chondrogenesis of human mesenchymal stem cells: effect of serum and medium supplements. Apoptosis 15:439-449. https://doi.org/10.1007/s10495-009-0431-x
  22. Yuan X, Sun Q, Ou Y, Wang S, Zhang W, Deng H, Wu X, Zhang L. 2014. Apoptosis is an obstacle to the differentiation of adipose-derived stromal cells into astrocytes. Neural. Regen. Res. 9:837-844. https://doi.org/10.4103/1673-5374.131600