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Home / Equine Research Bank / Stem Cells Cloning / Bone marrow mesenchymal stem cells as nuclear donors improve...
Stem cells and cloning : advances and applications2018; 11; 13-22; doi: 10.2147/SCCAA.S151763

Bone marrow mesenchymal stem cells as nuclear donors improve viability and health of cloned horses.

Abstract: Cell plasticity is crucial in cloning to allow an efficient nuclear reprogramming and healthy offspring. Hence, cells with high plasticity, such as multipotent mesenchymal stem cells (MSCs), may be a promising alternative for horse cloning. In this study, we evaluated the use of bone marrow-MSCs (BM-MSCs) as nuclear donors in horse cloning, and we compared the in vitro and in vivo embryo development with respect to fibroblasts. Methods: Zona-free nuclear transfer was performed using BM-MSCs (MSC group, n=3432) or adult fibroblasts (AF group, n=4527). Embryos produced by artificial insemination (AI) recovered by uterine flushing and transferred to recipient mares were used as controls (AI group). Results: Blastocyst development was higher in the MSC group than in the AF group (18.1% vs 10.9%, respectively; p<0.05). However, pregnancy rates and delivery rates were similar in both cloning groups, although they were lower than in the AI group (pregnancy rates: 17.7% [41/232] for MSC, 12.5% [37/297] for AF and 80.7% [71/88] for AI; delivery rates: 56.8% [21/37], 41.5% [17/41] and 90.1% [64/71], respectively). Remarkably, the gestation length of the AF group was significantly longer than the control (361.7±10.9 vs 333.9±8.7 days), in contrast to the MSC group (340.6±8.89 days). Of the total deliveries, 95.2% (20/21) of the MSC-foals were viable, compared to 52.9% (9/17) of the AF-foals (p<0.05). In addition, the AF-foals had more physiological abnormalities at birth than the MSC-foals; 90.5% (19/21) of the MSC-delivered foals were completely normal and healthy, compared to 35.3% (6/17) in the AF group. The abnormalities included flexural or angular limb deformities, umbilical cord enlargement, placental alterations and signs of syndrome of neonatal maladjustment, which were treated in most cases. Conclusions: In summary, we obtained 29 viable cloned foals and found that MSCs are suitable donor cells in horse cloning. Even more, these cells could be more efficiently reprogrammed compared to fibroblasts.
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Publication Date: 2018-02-14 PubMed ID: 29497320PubMed Central: PMC5818860DOI: 10.2147/SCCAA.S151763Google Scholar: Lookup
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Summary

This research summary has been generated with artificial intelligence and may contain errors and omissions. Refer to the original study to confirm details provided. Submit correction.

The research aims to identify if bone marrow-derived mesenchymal stem cells can be used as nuclear donors for horse cloning to improve the overall health and viability of cloned horses, as compared to the conventional method of using adult fibroblasts.

Research Methodology

  • The study conducted zona-free nuclear transfer with bone marrow-mesenchymal stem cells (grouped as MSC) and adult fibroblasts (grouped as AF).
  • A total of 3432 embryos were created using MSC and 4527 with AF.
  • The embryos created via artificial insemination and transferred to recipient mares served as the control group (grouped as AI).

Key Findings

  • The rate of blastocyst development (an early stage in embryo development) was significantly higher in the MSC group compared to the AF group.
  • While the rate of pregnancies and successful deliveries were more or less similar in MSC and AF groups, these rates were significantly lower compared to the AI group. Hence, the natural process of conception resulted in high success rates.
  • The gestational period for the AF group was significantly longer than both the control and the MSC group.
  • Among the deliveries, 95.2% of the cloned foals from the MSC group were viable compared to only 52.9% from the AF group. Health abnormalities such as limb deformities, umbilical cord enlargement, placental alterations and signs of neonatal maladjustment syndrome were more frequently observed in the AF group.

Conclusion

  • The experiments resulted in 29 viable cloned foals and demonstrated the suitability of MSCs as nuclear donor cells for horse cloning.
  • Given MSC’s higher plasticity, these cells can be reprogrammed more efficiently than fibroblasts.

Overall, the study clearly suggests that the use of MSCs can considerably improve the health and viability of cloned horses. These findings open new avenues in the field of cloning, suggesting potential alternatives to adult fibroblasts as nuclear donors.

Cite This Article

APA
Olivera R, Moro LN, Jordan R, Pallarols N, Guglielminetti A, Luzzani C, Miriuka SG, Vichera G. (2018). Bone marrow mesenchymal stem cells as nuclear donors improve viability and health of cloned horses. Stem Cells Cloning, 11, 13-22. https://doi.org/10.2147/SCCAA.S151763

Publication

Stem cells and cloning : advances and applications
ISSN: 1178-6957
NlmUniqueID: 101535817
Country: New Zealand
Language: English
Volume: 11
Pages: 13-22

Researcher Affiliations

Olivera, R
  • KHEIRON S.A Laboratory, Pilar, Buenos Aires, Argentina.
Moro, L N
  • LIAN-Unit Associated with CONICET, FLENI, Belen de Escobar, Buenos Aires, Argentina.
Jordan, R
  • KHEIRON S.A Laboratory, Pilar, Buenos Aires, Argentina.
Pallarols, N
  • Kawell Equine Hospital, Solís, Buenos Aires, Argentina.
Guglielminetti, A
  • Kawell Equine Hospital, Solís, Buenos Aires, Argentina.
Luzzani, C
  • LIAN-Unit Associated with CONICET, FLENI, Belen de Escobar, Buenos Aires, Argentina.
Miriuka, S G
  • LIAN-Unit Associated with CONICET, FLENI, Belen de Escobar, Buenos Aires, Argentina.
Vichera, G
  • KHEIRON S.A Laboratory, Pilar, Buenos Aires, Argentina.

Conflict of Interest Statement

Disclosure The authors report no conflicts of interest in this work.

References

This article includes 56 references
  1. Niemann H. Epigenetic reprogramming in mammalian species after SCNT-based cloning. Theriogenology 2016;86(1):80–90.
    pubmed: 27160443
  2. Waghmare SK, Estrada J, Reyes L. Gene targeting and cloning in pigs using fetal liver derived cells. J Surg Res 2011;171(2):e223–e229.
    pubmed: 21962810
  3. Richter A, Kurome M, Kessler B. Potential of primary kidney cells for somatic cell nuclear transfer mediated transgenesis in pig. BMC Biotechnol 2012;12:84.
    pmc: PMC3537537pubmed: 23140586
  4. Polejaeva IA, Chen SH, Vaught TD. Cloned pigs produced by nuclear transfer from adult somatic cells. Nature 2000;407(6800):86–90.
    pubmed: 10993078
  5. Lagutina I, Lazzari G, Duchi R. Somatic cell nuclear transfer in horses: effect of oocyte morphology, embryo reconstruction method and donor cell type. Reproduction 2005;130(4):559–567.
    pubmed: 16183874
  6. Jin HF, Kumar BM, Kim JG. Enhanced development of porcine embryos cloned from bone marrow mesenchymal stem cells. Int J dev Biol 2007;51(1):85–90.
    pubmed: 17183468
  7. Li Z, He X, Chen L. Bone marrow mesenchymal stem cells are an attractive donor cell type for production of cloned pigs as well as genetically modified cloned pigs by somatic cell nuclear transfer. Cell Reprogram 2013;15(5):459–470.
    pmc: PMC3787326pubmed: 24033142
  8. Lee JH, Lee WJ, Jeon RH. Development and gene expression of porcine cloned embryos derived from bone marrow stem cells with overexpressing Oct4 and Sox2. Cell Reprogram 2014;16(6):428–438.
    pmc: PMC4245852pubmed: 25437870
  9. Samiec M, Opiela J, LipiNski D, Romanek J. Trichostatin A-mediated epigenetic transformation of adult bone marrow-derived mesenchymal stem cells biases the in vitro developmental capability, quality, and pluripotency extent of porcine cloned embryos. Biomed Res Int 2015;2015:814686.
    pmc: PMC4381569pubmed: 25866813
  10. Song Z, Cong P, Ji Q. Establishment, differentiation, electroporation and nuclear transfer of porcine mesenchymal stem cells. Reprod Domest Anim 2015;50(5):840–148.
    pubmed: 26331974
  11. Yang Z, Vajta G, Xu Y. Production of pigs by hand-made cloning using mesenchymal stem cells and fibroblasts. Cell Reprogram 2016;18(4):256–263.
    pubmed: 27459584
  12. Nazari H, Shirazi A, Shams-Esfandabadi N, Aafzali A, Aahmadi E. The effect of amniotic membrane stem cells as donor nucleus on gene expression in reconstructed bovine oocytes. Int J Dev Biol 2016;60(4–6):95–102.
    pubmed: 27389982
  13. Da Silva CG, Martins CF, Cardoso TC. Production of bovine embryos and calves cloned by nuclear transfer using mesenchymal stem cells from amniotic fluid and adipose tissue. Cell Reprogram 2016;18(2):127–136.
    pubmed: 27055630
  14. Kwong PJ, Nam HY, Wan Khadijah WE, Kamarul T, Abdullah RB. Comparison of in vitro developmental competence of cloned caprine embryos using donor karyoplasts from adult bone marrow mesenchymal stem cells vs ear fibroblast cells. Reprod Domest Anim 2014;49(2):249–253.
    pubmed: 24456113
  15. Su X, Ling Y, Liu C. Isolation, culture, differentiation, and nuclear reprogramming of mongolian sheep fetal bone marrow-derived mesenchymal stem cells. Cell Reprogram 2015;17(4):288–296.
    pubmed: 26086202
  16. Olivera R, Moro LN, Jordan R. In vitro and in vivo development of horse cloned embryos generated with ipscs, mesenchymal stromal cells and fetal or adult fibroblasts as nuclear donors. PLoS One 2016;11(10):e0164049.
    pmc: PMC5061425pubmed: 27732616
  17. Vidal MA, Kilroy GE, Johnson JR, Lopez MJ, Moore RM, Gimble JM. Cell growth characteristics and differentiation frequency of adherent equine bone marrow-derived mesenchymal stromal cells: adipogenic and osteogenic capacity. Vet Surg 2006;35(7):601–610.
    pubmed: 17026544
  18. Arnhold SJ, Goletz I, Klein H. Isolation and characterization of bone marrow-derived equine mesenchymal stem cells. Am J Vet Res 2007;68(10):1095–1105.
    pubmed: 17916017
  19. Koch TG, Heerkens T, Thomsen PD, Betts DH. Isolation of mesenchymal stem cells from equine umbilical cord blood. BMC Biotechnol 2007;7:26.
    pmc: PMC1904213pubmed: 17537254
  20. Reed SA, Johnson SE. Equine umbilical cord blood contains a population of stem cells that express Oct 4 and differentiate into mesodermal and endodermal cell types. J Cell Physiol 2008;215(2):329–336.
    pubmed: 17929245
  21. Hoynowski SM, Fry MM, Gardner BM. Characterization and differentiation of equine umbilical cord-derived matrix cells. Biochem Biophys Res Commun 2007;362(2):347–353.
    pubmed: 17719011
  22. Vidal MA, Kilroy GE, Lopez MJ, Johnson JR, Moore RM, Gimble JM. Characterization of equine adipose tissue-derived stromal cells: Adipogenic and osteogenic capacity and comparison with bone marrow-derived mesenchymal stromal cells. Vet Surg 2007;36(7):613–622.
    pubmed: 17894587
  23. Koerner J, Nesic D, Romero JD, Brehm W, Mainil-Varlet P, Grogan SP. Equine peripheral blood-derived progenitors in comparison to bone marrow-derived mesenchymal stem cells. Stem Cells 2006;24(6):1613–1619.
    pubmed: 16769763
  24. Smith RKW, Korda M, Blunn GW, Goodship AE. Isolation and implantation of autologous equine mesenchymal stem cells from bone marrow into the superficial digital flexor tendon as a potential novel treatment. Equine Vet J 2003;35(1):99–102.
    pubmed: 12553472
  25. Guest DJ, Ousey JC, Smith MR. Defining the expression of marker genes in equine mesenchymal stromal cells. Stem Cells Cloning 2008;1:1–9.
    pmc: PMC3781685pubmed: 24198500
  26. Wilke MM, Nydam DV, Nixon AJ. Enhanced early chondrogenesis in articular defects following arthroscopic mesenchymal stem cell implantation in an equine model. J Orthop Res 2007;25(7):913–925.
    pubmed: 17405160
  27. Lepage SI, Nagy K, Sung HK, Kandel RA, Nagy A, Koch TG. Generation, characterization, and multilineage potency of mesenchymal-like progenitors derived from equine induced pluripotent stem cells. Stem Cells Dev 2016;25(1):80–89.
    pubmed: 26414480
  28. Esteves CL, Sharma R, Dawson L. Expression of putative markers of pluripotency in equine embryonic and adult tissues. Vet J 2014;202(3):533–535.
    pubmed: 25241949
  29. Lagutina I, Lazzari G, Duchi R. Comparative aspects of somatic cell nuclear transfer with conventional and zona-free method in cattle, horse, pig and sheep. Theriogenology 2007;67(1):90–98.
    pubmed: 17081599
  30. Gambini A, Jarazo J, Olivera R, Salamone DF. Equine cloning: in vitro and in vivo development of aggregated embryos. Biol Reprod 2012;87(1):1–9.
    pubmed: 22553223
  31. Gambini A, De Stefano A, Bevacqua RJ, Karlanian F, Salamone DF. The aggregation of four reconstructed zygotes is the limit to improve the developmental competence of cloned equine embryos. PLoS One 2014;9:e110998.
    pmc: PMC4232247pubmed: 25396418
  32. Choi YH, Velez IC, Macías-García B, Hinrichs K. Timing factors affecting blastocyst development in equine somatic cell nuclear transfer. Cell Reprogram 2015;17(2):124–130.
    pubmed: 25826725
  33. Johnson AK, Clark-Price SC, Choi YH, Hartman DL, Hinrichs K. Physical and clinicopathologic findings in foals derived by use of somatic cell nuclear transfer: 14 cases (2004±2008). J Am Vet Med Assoc 2010;236(9):983–990.
    pubmed: 20433399
  34. Choi YH, Norris JD, Velez IC, Jacobson CC, Hartman DL, Hinrichs K. A viable foal obtained by equine somatic cell nuclear transfer using oocytes recovered from immature follicles of live mares. Theriogenology 2013;79(5):791–796.
    pubmed: 23312717
  35. Hall V, Hinrichs K, Lazzari G, Betts DH, Hyttel P. Early embryonic development, assisted reproductive technologies, and pluripotent stem cell biology in domestic mammals. Vet J 2013;197(2):128–142.
    pubmed: 23810186
  36. Bavister B, Yanagimachi R. The effects of sperm extracts and energy sources on the motility and acrosome reaction of hamster spermatozoa in vitro. Biol Reprod 1977;16(2):228–237.
    pubmed: 831847
  37. Galli C, Lagutina I, Crotti G. Pregnancy: a cloned horse born to its dam twin. Nature 2003;424(6949):635.
    pubmed: 12904778
  38. Kumar BM, Jin HF, Kim JG. Differential gene expression patterns in porcine nuclear transfer embryos reconstructed with fetal fibroblasts and mesenchymal stem cells. Dev Dyn 2007;236(2):435–446.
    pubmed: 17191234
  39. Woods GL, White KL, Vanderwall DK. A mule cloned from fetal cells by nuclear transfer. Science 2003;301(5636):1063.
    pubmed: 12775846
  40. Vanderwall DK, Woods GL, Aston KI. Cloned horse pregnancies produced using adult cumulus cells. Reprod Fertil Dev 2004;16(7):675–679.
    pubmed: 15740690
  41. Wonyou L, Kilyoung S, Inhyung L, Hyungdo S, Byeong CL, Seongchan Y. Cloned foal derived from in vivo matured horse oocytes aspirated by the short disposable needle system. J Vet Sci 2015;16(4):509–516.
    pmc: PMC4701744pubmed: 26119166
  42. Cummins C, Carrington S, Fitzpatrick E, Duggan V. Ascending placentitis in the mare: a review. Ir Vet J 2008;61(5):307–313.
    pmc: PMC3113861pubmed: 21851713
  43. Schmidt M, Kragh PM, Li J. Pregnancies and piglets from large white sow recipients after two transfer methods of cloned and transgenic embryos of different pig breeds. Theriogenology 2010;74(7):1233–1240.
    pubmed: 20688371
  44. Heyman Y, Chavatte-Palmer P, LeBourhis D, Camous S, Vignon X, Renard JP. Frequency and occurrence of late-gestation losses from cattle cloned embryos. Biol Reprod 2002;66(1):6–13.
    pubmed: 11751257
  45. Berg D, Li C, Asher G, Wells D, Oback B. Red deer cloned from antler stem cells and their differentiated progeny. Biol Reprod 2007;77(3):384–394.
    pubmed: 17522075
  46. Oback B. Cloning from stem cells: different lineages, different species, same story. Reprod Fertil Dev 2009;21(1):83–94.
    pubmed: 19152749
  47. Secher J, Liu Y, Petkov S. Evaluation of porcine stem cell competence for somatic cell nuclear transfer and production of cloned animals. Anim Reprod Sci 2017;178:40–49.
    pubmed: 28126267
  48. Faast R, Harrison S, Beebe L, McIlfatrick S, Ashman R, Nottle M. Use of adult mesenchymal stem cells isolated from bone marrow and blood for somatic cell nuclear transfer in pigs. Cloning Stem Cells 2006;8(3):166–173.
    pubmed: 17009893
  49. Fan N, Chen J, Shang Z. Piglets cloned from induced pluripotent stem cells. Cell Res 2013;23(1):162–166.
    pmc: PMC3541650pubmed: 23247628
  50. Du X, Feng T, Yu D. Barriers for deriving transgene-Free pig iPS cells with episomal vectors. Stem Cells 2015;33(11):3228–3238.
    pmc: PMC5025037pubmed: 26138940
  51. Takahashi K, Yamanaka S. Induction of pluripotent stem cells from mouse embryonic and adult fibroblast cultures by defined factors. Cell 2006;126(4):663–676.
    pubmed: 16904174
  52. Hall V, Hyttel P. Breaking down pluripotency in the porcine embryo reveals both a premature and reticent stem cell state in the inner cell mass and unique expression profiles of the naive and primed stem cell states. Stem Cells Dev 2014;23(17):2030–2045.
    pubmed: 24742229
  53. Yuan Y, Lee K, Park K. Cell cycle synchronization of leukemia inhibitory factor (LIF)-dependent porcine-induced pluripotent stem cells and the generation of cloned embryos. ABBV Cell Cycle 2014;13(18):1265–1276.
    pmc: PMC4049963pubmed: 24621508
  54. Smith RK, Werling NJ, Dakin SG, Alam R, Goodship AE, Dudhia J. Beneficial effects of autologous bone marrow-derived mesenchymal stem cells in naturally occurring tendinopathy. PLoS One 2013;8(9):e75697.
    pmc: PMC3783421pubmed: 24086616
  55. Broeckx S, Zimmerman M, Crocetti S. Regenerative therapies for equine degenerative joint disease: a preliminary study. PLoS One 2014;9(1):e85917.
    pmc: PMC3896436pubmed: 24465787
  56. Geburek F, Roggel F, van Schie HTM. Effect of single intralesional treatment of surgically induced equine superficial digital flexor tendon core lesions with adipose-derived mesenchymal stromal cells: a controlled experimental trial. Stem Cell Res Ther 2017;8(1):129.
    pmc: PMC5460527pubmed: 28583184

Citations

This article has been cited 11 times.
  1. Lee SC, Lee WJ, Son YB, Jin YB, Lee HJ, Bok E, Lee S, Lee SY, Jo CH, Kim TS, Hong CY, Kang SY, Rho GJ, Choe YH, Lee SL. Trichostatin A-Induced Epigenetic Modifications and Their Influence on the Development of Porcine Cloned Embryos Derived from Bone Marrow-Mesenchymal Stem Cells. Int J Mol Sci 2025 Mar 6;26(5).
    doi: 10.3390/ijms26052359pubmed: 40076980google scholar: lookup
  2. Samiec M. Molecular Mechanisms of Somatic Cell Cloning and Other Assisted Reproductive Technologies in Mammals: Which Determinants Have Been Unraveled Thus Far?-Current Status, Further Progress and Future Challenges. Int J Mol Sci 2024 Dec 21;25(24).
    doi: 10.3390/ijms252413675pubmed: 39769437google scholar: lookup
  3. Wu Y, Wang C, Fan X, Ma Y, Liu Z, Ye X, Shen C, Wu C. The impact of induced pluripotent stem cells in animal conservation. Vet Res Commun 2024 Apr;48(2):649-663.
    doi: 10.1007/s11259-024-10294-3pubmed: 38228922google scholar: lookup
  4. Suvá M, Arnold VH, Wiedenmann EA, Jordan R, Galvagno E, Martínez M, Vichera GD. First sex modification case in equine cloning. PLoS One 2023;18(1):e0279869.
    doi: 10.1371/journal.pone.0279869pubmed: 36598913google scholar: lookup
  5. Wang DH, Wu XM, Chen JS, Cai ZG, An JH, Zhang MY, Li Y, Li FP, Hou R, Liu YL. Isolation and characterization mesenchymal stem cells from red panda (Ailurus fulgens styani) endometrium. Conserv Physiol 2022 Jan 1;10(1):coac004.
    doi: 10.1093/conphys/coac004pubmed: 35211318google scholar: lookup
  6. Antczak DF, Allen WRT. Placentation in Equids. Adv Anat Embryol Cell Biol 2021;234:91-128.
    doi: 10.1007/978-3-030-77360-1_6pubmed: 34694479google scholar: lookup
  7. Jin X, Hao Z, Zhao M, Shen J, Ke N, Song Y, Qiao L, Lu Y, Hu L, Wu X, Wang J, Luo Y. MicroRNA-148a Regulates the Proliferation and Differentiation of Ovine Preadipocytes by Targeting PTEN. Animals (Basel) 2021 Mar 15;11(3).
    doi: 10.3390/ani11030820pubmed: 33803986google scholar: lookup
  8. Samiec M, Skrzyszowska M. Extranuclear Inheritance of Mitochondrial Genome and Epigenetic Reprogrammability of Chromosomal Telomeres in Somatic Cell Cloning of Mammals. Int J Mol Sci 2021 Mar 18;22(6).
    doi: 10.3390/ijms22063099pubmed: 33803567google scholar: lookup
  9. Hisey EA, Ross PJ, Meyers S. Genetic Manipulation of the Equine Oocyte and Embryo. J Equine Vet Sci 2021 Apr;99:103394.
    doi: 10.1016/j.jevs.2021.103394pubmed: 33781418google scholar: lookup
  10. Moro LN, Viale DL, Bastón JI, Arnold V, Suvá M, Wiedenmann E, Olguín M, Miriuka S, Vichera G. Generation of myostatin edited horse embryos using CRISPR/Cas9 technology and somatic cell nuclear transfer. Sci Rep 2020 Sep 24;10(1):15587.
    doi: 10.1038/s41598-020-72040-4pubmed: 32973188google scholar: lookup
  11. Moro LN, Amin G, Furmento V, Waisman A, Garate X, Neiman G, La Greca A, Santín Velazque NL, Luzzani C, Sevlever GE, Vichera G, Miriuka SG. MicroRNA characterization in equine induced pluripotent stem cells. PLoS One 2018;13(12):e0207074.
    doi: 10.1371/journal.pone.0207074pubmed: 30507934google scholar: lookup
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