Journal of Veterinary Science

The Korean Society of Veterinary Science (대한수의학회)
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Evaluation of porcine urine-derived cells as nuclei donor for somatic cell nuclear transfer

  • Zhang, Yu-Ting (Key Laboratory of Animal Cellular and Genetics Engineering of Heilongjiang Province, College of Life Science, Northeast Agricultural University) ;
  • Yao, Wang (Key Laboratory of Animal Cellular and Genetics Engineering of Heilongjiang Province, College of Life Science, Northeast Agricultural University) ;
  • Chai, Meng-Jia (Key Laboratory of Animal Cellular and Genetics Engineering of Heilongjiang Province, College of Life Science, Northeast Agricultural University) ;
  • Liu, Wen-Jing (Key Laboratory of Animal Cellular and Genetics Engineering of Heilongjiang Province, College of Life Science, Northeast Agricultural University) ;
  • Liu, Yan (Key Laboratory of Animal Cellular and Genetics Engineering of Heilongjiang Province, College of Life Science, Northeast Agricultural University) ;
  • Liu, Zhong-Hua (Key Laboratory of Animal Cellular and Genetics Engineering of Heilongjiang Province, College of Life Science, Northeast Agricultural University) ;
  • Weng, Xiao-Gang (Key Laboratory of Animal Cellular and Genetics Engineering of Heilongjiang Province, College of Life Science, Northeast Agricultural University)
  • Received : 2021.12.07
  • Accepted : 2022.01.20
  • Published : 2022.03.31
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Abstract

Background: Somatic cell nuclear transfer (SCNT) is used widely in cloning, stem cell research, and regenerative medicine. The type of donor cells is a key factor affecting the SCNT efficiency. Objectives: This study examined whether urine-derived somatic cells could be used as donors for SCNT in pigs. Methods: The viability of cells isolated from urine was assessed using trypan blue and propidium iodide staining. The H3K9me3/H3K27me3 level of the cells was analyzed by immunofluorescence. The in vitro developmental ability of SCNT embryos was evaluated by the blastocyst rate and the expression levels of the core pluripotency factor. Blastocyst cell apoptosis was examined using a terminal deoxynucleotidyl transferase dUTP nick end-labeling assay. The in vivo developmental ability of SCNT embryos was evaluated after embryo transfer. Results: Most sow urine-derived cells were viable and could be cultured and propagated easily. On the other hand, most of the somatic cells isolated from the boar urine exhibited poor cellular activity. The in vitro development efficiency between the embryos produced by SCNT using porcine embryonic fibroblasts (PEFs) and urine-derived cells were similar. Moreover, The H3K9me3 in SCNT embryos produced from sow urine-derived cells and PEFs at the four-cell stage showed similar intensity. The levels of Oct4, Nanog, and Sox2 expression in blastocysts were similar in the two groups. Furthermore, there is a similar apoptotic level of cloned embryos produced by the two types of cells. Finally, the full-term development ability of the cloned embryos was evaluated, and the cloned fetuses from the urine-derived cells showed absorption. Conclusions: Sow urine-derived cells could be used to produce SCNT embryos.

Keywords

Acknowledgement

This study was supported by the Natural Science Foundation of Heilongjiang Province (grant number C2017035); and the Postdoctoral Science Foundation of Heilongjiang Province (grant number LBH-Z17010).

References

  1. Wilmut I, Schnieke AE, McWhir J, Kind AJ, Campbell KH. Viable offspring derived from fetal and adult mammalian cells. Nature. 1997;385(6619):810-813. https://doi.org/10.1038/385810a0
  2. Kato M, Han TW, Xie S, Shi K, Du X, Wu LC, et al. Cell-free formation of RNA granules: low complexity sequence domains form dynamic fibers within hydrogels. Cell. 2012;149(4):753-767. https://doi.org/10.1016/j.cell.2012.04.017
  3. Matoba S, Zhang Y. Somatic cell nuclear transfer reprogramming: mechanisms and applications. Cell Stem Cell. 2018;23(4):471-485. https://doi.org/10.1016/j.stem.2018.06.018
  4. Kishi M, Itagaki Y, Takakura R, Imamura M, Sudo T, Yoshinari M, et al. Nuclear transfer in cattle using colostrum-derived mammary gland epithelial cells and ear-derived fibroblast cells. Theriogenology. 2000;54(5):675-684. https://doi.org/10.1016/S0093-691X(00)00382-4
  5. Nel-Themaat L, Gomez MC, Damiani P, Wirtu G, Dresser BL, Bondioli KR, et al. Isolation, culture and characterisation of somatic cells derived from semen and milk of endangered sheep and eland antelope. Reprod Fertil Dev. 2007;19(4):576-584. https://doi.org/10.1071/RD06153
  6. Hintz DS, Sens MA, Jenkins MQ, Sens DA. Tissue culture of epithelial cells from urine. I. Serum-free growth of cells from newborn infants. Pediatr Pathol. 1984;2(2):153-163. https://doi.org/10.3109/15513818409025883
  7. Uhm KO, Jo EH, Go GY, Kim SJ, Choi HY, Im YS, et al. Generation of human induced pluripotent stem cells from urinary cells of a healthy donor using a non-integration system. Stem Cell Res (Amst). 2017;21:44-46. https://doi.org/10.1016/j.scr.2017.03.019
  8. Zhou T, Benda C, Duzinger S, Huang Y, Li X, Li Y, et al. Generation of induced pluripotent stem cells from urine. J Am Soc Nephrol. 2011;22(7):1221-1228. https://doi.org/10.1681/ASN.2011010106
  9. Zhou T, Benda C, Dunzinger S, Huang Y, Ho JC, Yang J, et al. Generation of human induced pluripotent stem cells from urine samples. Nat Protoc. 2012;7(12):2080-2089. https://doi.org/10.1038/nprot.2012.115
  10. Cheng L, Lei Q, Yin C, Wang HY, Jin K, Xiang M. Generation of urine cell-derived non-integrative human iPSCs and iNSCs: a step-by-step optimized protocol. Front Mol Neurosci. 2017;10:348. https://doi.org/10.3389/fnmol.2017.00348
  11. Kloskowski T, Nowacki M, Pokrywczynska M, Drewa T. Urine--a waste or the future of regenerative medicine? Med Hypotheses. 2015;84(4):344-349. https://doi.org/10.1016/j.mehy.2015.01.019
  12. Mizutani E, Torikai K, Wakayama S, Nagatomo H, Ohinata Y, Kishigami S, et al. Generation of cloned mice and nuclear transfer embryonic stem cell lines from urine-derived cells. Sci Rep. 2016;6(1):23808. https://doi.org/10.1038/srep23808
  13. Madheshiya PK, Sahare AA, Jyotsana B, Singh KP, Saini M, Raja AK, et al. Production of a cloned buffalo (Bubalus bubalis) calf from somatic cells isolated from urine. Cell Reprogram. 2015;17(3):160-169. https://doi.org/10.1089/cell.2014.0097
  14. Liu Z, Song J, Wang Z, Tian J, Kong Q, Zheng Z, et al. Green fluorescent protein (GFP) transgenic pig produced by somatic cell nuclear transfer. Chin Sci Bull. 2008;53(7):1035-1039.
  15. Xie B, Zhang H, Wei R, Li Q, Weng X, Kong Q, et al. Histone H3 lysine 27 trimethylation acts as an epigenetic barrier in porcine nuclear reprogramming. Reproduction. 2016;151(1):9-16. https://doi.org/10.1530/REP-15-0338
  16. He W, Zhu W, Cao Q, Shen Y, Zhou Q, Yu P, et al. Generation of mesenchymal-like stem cells from urine in pediatric patients. Transplant Proc. 2016;48(6):2181-2185. https://doi.org/10.1016/j.transproceed.2016.02.078
  17. Skoberne A, Konieczny A, Schiffer M. Glomerular epithelial cells in the urine: what has to be done to make them worthwhile? Am J Physiol Renal Physiol. 2009;296(2):F230-F241. https://doi.org/10.1152/ajprenal.90507.2008
  18. Loh EH, Keng VW, Ward PB. Blood cells and red cell morphology in the urine of healthy children. Clin Nephrol 1990;34(4):185-187.
  19. Sutherland GR, Bain AD. Culture of cells from the urine of newborn children. Nature. 1972;239(5369):231. https://doi.org/10.1038/239231a0
  20. Braude H, Forfar JO, Gould JC, McLeod JW. Cell and bacterial counts in the urine of normal infants and children. BMJ. 1967;4(5581):697-701. https://doi.org/10.1136/bmj.4.5581.697
  21. Bharadwaj S, Liu G, Shi Y, Wu R, Yang B, He T, et al. Multipotential differentiation of human urine-derived stem cells: potential for therapeutic applications in urology. Stem Cells. 2013;31(9):1840-1856. https://doi.org/10.1002/stem.1424
  22. Dmitrieva NI, Michea LF, Rocha GM, Burg MB. Cell cycle delay and apoptosis in response to osmotic stress. Comp Biochem Physiol A Mol Integr Physiol. 2001;130(3):411-420. https://doi.org/10.1016/S1095-6433(01)00439-1
  23. Burg MB, Ferraris JD, Dmitrieva NI. Cellular response to hyperosmotic stresses. Physiol Rev. 2007;87(4):1441-1474. https://doi.org/10.1152/physrev.00056.2006
  24. Wakayama S, Ohta H, Hikichi T, Mizutani E, Iwaki T, Kanagawa O, et al. Production of healthy cloned mice from bodies frozen at -20 degrees C for 16 years. Proc Natl Acad Sci U S A. 2008;105(45):17318-17322. https://doi.org/10.1073/pnas.0806166105
  25. Wakayama S, Ohta H, Kishigami S, Thuan NV, Hikichi T, Mizutani E, et al. Establishment of male and female nuclear transfer embryonic stem cell lines from different mouse strains and tissues. Biol Reprod. 2005;72(4):932-936. https://doi.org/10.1095/biolreprod.104.035105
  26. Adamowicz J, Kloskowski T, Tworkiewicz J, Pokrywczynska M, Drewa T. Urine is a highly cytotoxic agent: does it influence stem cell therapies in urology? Transplant Proc. 2012;44(5):1439-1441. https://doi.org/10.1016/j.transproceed.2012.01.128
  27. Matoba S, Liu Y, Lu F, Iwabuchi KA, Shen L, Inoue A, et al. Embryonic development following somatic cell nuclear transfer impeded by persisting histone methylation. Cell. 2014;159(4):884-895. https://doi.org/10.1016/j.cell.2014.09.055
  28. Chung YG, Matoba S, Liu Y, Eum JH, Lu F, Jiang W, et al. Histone demethylase expression enhances human somatic cell nuclear transfer efficiency and promotes derivation of pluripotent stem cells. Cell Stem Cell. 2015;17(6):758-766. https://doi.org/10.1016/j.stem.2015.10.001
  29. Xin L, Wang YZ, Gao YP, Su JM, Zhang JC, Xing XP, et al. H3K9 demethylase KDM4E is an epigenetic regulator for bovine embryonic development and a defective factor for nuclear reprogramming. Development. 2018;145(4):dev158261. https://doi.org/10.1242/dev.158261
  30. Ruan D, Peng J, Wang X, Ouyang Z, Zou Q, Yang Y, et al. XIST Derepression in active X chromosome hinders pig somatic cell nuclear transfer. Stem Cell Reports. 2018;10(2):494-508. https://doi.org/10.1016/j.stemcr.2017.12.015
  31. Huang Y, Ouyang H, Yu H, Lai L, Pang D, Li Z. Efficiency of porcine somatic cell nuclear transfer - a retrospective study of factors related to embryo recipient and embryos transferred. Biol Open. 2013;2(11):1223-1228. https://doi.org/10.1242/bio.20135983