Journal of Animal Reproduction and Biotechnology (한국동물생명공학회지)

The Korean Society of Animal Reproduction and Biotechnology (한국동물생명공학회)
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Porcine somatic cell nuclear transfer using telomerase reverse transcriptase-transfected mesenchymal stem cells reduces apoptosis induced by replicative senescence

  • Jeon, Ryounghoon (College of Veterinary Medicine, Gyeongsang National University) ;
  • Rho, Gyu-Jin (College of Veterinary Medicine, Gyeongsang National University)
  • Received : 2020.06.15
  • Accepted : 2020.08.04
  • Published : 2020.09.30
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Abstract

Mesenchymal stem cells (MSCs) have been widely used as donor cells for somatic cell nuclear transfer (SCNT) to increase the efficiency of embryo cloning. Since replicative senescence reduces the efficiency of embryo cloning in MSCs during in vitro expansion, transfection of telomerase reverse transcriptase (TERT) into MSCs has been used to suppress the replicative senescence. Here, TERT-transfected MSCs in comparison with early passage MSCs (eMSCs) and sham-transfected MSCs (sMSCs) were used to evaluate the effects of embryo cloning with SCNT in a porcine model. Cloned embryos from tMSC, eMSC, and sMSC groups were indistinguishable in their fusion rate, cleavage rate, total cell number, and gene expression levels of OCT4, SOX2 and NANOG during the blastocyst stage. The blastocyst formation rates of tMSC and sMSC groups were comparable but significantly lower than that of the eMSC group (p < 0.05). In contrast, tMSC and eMSC groups demonstrated significantly reduced apoptotic incidence (p < 0.05), and decreased BAX but increased BCL2 expression in the blastocyst stage compared to the sMSC group (p < 0.05). Therefore, MSCs transfected with telomerase reverse transcriptase do not affect the overall development of the cloned embryos in porcine SCNT, but enables to maintain embryo quality, similar to apoptotic events in SCNT embryos typically achieved by an early passage MSC. This finding offers a bioengineering strategy in improving the porcine cloned embryo quality.

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References

  1. Daniel SM, Raipuria P, Sarkhel BC. 2008. Efficiency of cloned embryo production using different types of cell donor and electric fusion strengths in goats. Small Rumin. Res. 77:45-50. https://doi.org/10.1016/j.smallrumres.2008.02.001
  2. Del Bufalo D, Rizzo A, Trisciuoglio D, Cardinali G, Torrisi MR, Zangemeister-Wittke U, Zupi G, Biroccio A. 2005. Involvement of hTERT in apoptosis induced by interference with Bcl-2 expression and function. Cell Death Differ. 12:1429-1438. https://doi.org/10.1038/sj.cdd.4401670
  3. Fabian D, Koppel J, Maddox-Hyttel P. 2005. Apoptotic processes during mammalian preimplantation development. Theriogenology 64:221-231. https://doi.org/10.1016/j.theriogenology.2004.11.022
  4. Gouveia C, Huyser C, Egli D, Pepper MS. 2020. Lessons learned from somatic cell nuclear transfer. Int. J. Mol. Sci. 21:2314. https://doi.org/10.3390/ijms21072314
  5. Hao Y, Lai L, Mao J, Im GS, Bonk A, Prather RS. 2003. Apoptosis and in vitro development of preimplantation porcine embryos derived in vitro or by nuclear transfer. Biol. Reprod. 69:501-507. https://doi.org/10.1095/biolreprod.103.016170
  6. Hiyama E and Hiyama K. 2007. Telomere and telomerase in stem cells. Br. J. Cancer 96:1020-1024. https://doi.org/10.1038/sj.bjc.6603671
  7. Jeon R, Park S, Lee SL, Rho GJ. 2020. Subpopulations of miniature pig mesenchymal stromal cells with different differentiation potentials differ in the expression of octamer-binding transcription factor 4 and sex determining region Y-box 2. Asian-Australas J. Anim. Sci. 33:515-524. https://doi.org/10.5713/ajas.19.0416
  8. Jeon RH, Maeng GH, Lee WJ, Kim TH, Lee YM, Lee JH, Kumar BM, Lee SL, Rho GJ. 2012. Removal of cumulus cells before oocyte nuclear maturation enhances enucleation rates without affecting the developmental competence of porcine cloned embryos. Jpn. J. Vet. Res. 60:191-203.
  9. Kanaya T, Kyo S, Hamada K, Takakura M, Kitagawa Y, Harada H, Inoue M. 2000. Adenoviral expression of p53 represses telomerase activity through down-regulation of human telomerase reverse transcriptase transcription. Clin. Cancer Res. 6:1239-1247.
  10. Kumar BM, Jin HF, Kim JG, Ock SA, Hong Y, Balasubramanian S, Choe SY, Rho GJ. 2007. Differential gene expression patterns in porcine nuclear transfer embryos reconstructed with fetal fibroblasts and mesenchymal stem cells. Dev. Dyn. 236:435-446. https://doi.org/10.1002/dvdy.21042
  11. Kumar BM, Maeng GH, Jeon RH, Lee YM, Lee WJ, Jeon BG, Ock SA, Rho GJ. 2012. Developmental expression of lineage specific genes in porcine embryos of different origins. J. Assist. Reprod. Genet. 29:723-733. https://doi.org/10.1007/s10815-012-9797-8
  12. Lee AR, Hong K, Choi SH, Park C, Park JK, Lee JI, Bang JI, Seol DW, Lee JE, Lee DR. 2019. Anti-apoptotic regulation contributes to the successful nuclear reprogramming using cryopreserved oocytes. Stem Cell Reports 12:545-556. https://doi.org/10.1016/j.stemcr.2019.01.019
  13. Lee JH, Lee WJ, Jeon RH, Lee YM, Jang SJ, Lee SL, Jeon BG, Ock SA, King WA, Rho GJ. 2014. Development and gene expression of porcine cloned embryos derived from bone marrow stem cells with overexpressing Oct4 and Sox2. Cell. Reprogram. 16:428-438. https://doi.org/10.1089/cell.2014.0036
  14. Lee WJ, Jang SJ, Lee SC, Park JS, Jeon RH, Subbarao RB, Bharti D, Shin JK, Park BW, Rho GJ. 2017. Selection of reference genes for quantitative real-time polymerase chain reaction in porcine embryos. Reprod. Fertil. Dev. 29:357-367. https://doi.org/10.1071/RD14393
  15. Legzdina D, Romanauska A, Nikulshin S, Kozlovska T, Berzins U. 2016. Characterization of senescence of culture-expanded Human adipose-derived mesenchymal stem cells. Int. J. Stem Cells 9:124-136. https://doi.org/10.15283/ijsc.2016.9.1.124
  16. Liu TM, Ng WM, Tan HS, Vinitha D, Yang Z, Fan JB, Zou Y, Hui JH, Lee EH, Lim B. 2013. Molecular basis of immortalization of human mesenchymal stem cells by combination of p53 knockdown and human telomerase reverse transcriptase overexpression. Stem Cells Dev. 22:268-278. https://doi.org/10.1089/scd.2012.0222
  17. Meissner A and Jaenisch R. 2006. Mammalian nuclear transfer. Dev. Dyn. 235:2460-2469. https://doi.org/10.1002/dvdy.20915
  18. Mulligan B, Hwang JY, Kim HM, Oh JN, Choi KH, Lee CK. 2012. Pro-apoptotic effect of pifithrin-${\alpha}$ on preimplantation porcine in vitro fertilized embryo development. Asian-Australas. J. Anim. Sci. 25:1681-1690. https://doi.org/10.5713/ajas.2012.12404
  19. Ock SA, Lee SL, Kim JG, Kumar BM, Balasubramanian S, Choe SY, Rho GJ. 2007. Development and quality of porcine embryos in different culture system and embryo-producing methods. Zygote 15:1-8. https://doi.org/10.1017/S0967199406003911
  20. Turinetto V, Vitale E, Giachino C. 2016. Senescence in human mesenchymal stem cells: functional changes and implications in stem cell-based therapy. Int. J. Mol. Sci. 17:1164. https://doi.org/10.3390/ijms17071164
  21. Yuan Y, Lee K, Park KW, Spate LD, Prather RS, Wells KD, Roberts RM. 2014. Cell cycle synchronization of leukemia inhibitory factor (LIF)-dependent porcine-induced pluripotent stem cells and the generation of cloned embryos. Cell Cycle 13:1265-1276. https://doi.org/10.4161/cc.28176
  22. Zaim M, Karaman S, Cetin G, Isik S. 2012. Donor age and long-term culture affect differentiation and proliferation of human bone marrow mesenchymal stem cells. Ann. Hematol. 91:1175-1186. https://doi.org/10.1007/s00277-012-1438-x