FREE SHIPPING over $40
OVER 9000 FIVE-STAR REVIEWS
5% OFF ALL SUBSCRIPTIONS
100% HAPPINESS GUARANTEE
Analyze Diet
0
Home / Equine Research Bank / Stem Cells Int / Generation and miRNA Characterization of Equine Induced Plur...
Stem cells international2019; 2019; 1393791; doi: 10.1155/2019/1393791

Generation and miRNA Characterization of Equine Induced Pluripotent Stem Cells Derived from Fetal and Adult Multipotent Tissues.

Abstract: Pluripotent stem cells are believed to have greater clinical potential than mesenchymal stem cells due to their ability to differentiate into almost any cell type of an organism, and since 2006, the generation of patient-specific induced pluripotent stem cells (iPSCs) has become possible in multiple species. Objective: We hypothesize that different cell types respond differently to the reprogramming process; thus, the goals of this study were to isolate and characterize equine adult and fetal cells and induce these cells to pluripotency for future regenerative and translational purposes. Methods: Adult equine fibroblasts (eFibros) and mesenchymal cells derived from the bone marrow (eBMmsc), adipose tissue (eADmsc), and umbilical cord tissue (eUCmsc) were isolated, their multipotency was characterized, and the cells were induced into pluripotency (eiPSCs). eiPSCs were generated through a lentiviral system using the factors OCT4, SOX2, c-MYC, and KLF4. The morphology and pluripotency maintenance potential (alkaline phosphatase detection, embryoid body formation, spontaneous differentiation, and expression of pluripotency markers) of the eiPSCs were characterized. Additionally, a miRNA profile analysis of the mesenchymal and eiPSCs was performed. Results: Multipotent cells were successfully isolated, but the eBMmsc failed to generate eiPSCs. The eADmsc-, eUCmsc-, and eFibros-derived iPSCs were positive for alkaline phosphatase, OCT4 and NANOG, were exclusively dependent on bFGF, and formed embryoid bodies. The miRNA profile revealed a segregated pattern between the eiPSCs and multipotent controls: the levels of miR-302/367 and the miR-92 family were increased in the eiPSCs, while the levels of miR-23, miR-27, and miR-30, as well as the let-7 family were increased in the nonpluripotent cells. Conclusions: We were able to generate bFGF-dependent iPSCs from eADmsc, eUCmsc, and eFibros with human OSKM, and the miRNA profile revealed that clonal lines may respond differently to the reprogramming process.
Get Full Text
Publication Date: 2019-05-02 PubMed ID: 31191664PubMed Central: PMC6525926DOI: 10.1155/2019/1393791Google Scholar: Lookup
The Equine Research Bank provides access to a large database of publicly available scientific literature. Inclusion in the Research Bank does not imply endorsement of study methods or findings by Mad Barn.
LinkedInCopy
  • Journal Article

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 explores the derivation and characterization of pluripotent stem cells (cells that can transform into any cell type) from a variety of adult and fetal equine tissues, with a particular focus on microRNA (miRNA) signatures. Interestingly, the researchers discover that different equine cell lines react differently to the reprogramming process to create these pluripotent stem cells.

Research Procedure

  • The researchers began by isolating adult equine fibroblasts and mesenchymal cells from various tissues – including bone marrow, adipose tissue, and umbilical cord tissue.
  • They confirmed these cells’ multipotency, meaning their ability to differentiate into several types of cells.
  • These cells were then induced to reach a state of pluripotency, transforming into what the team calls equine induced Pluripotent Stem Cells (eiPSCs).
  • The pluripotency was achieved using a lentiviral system with OCT4, SOX2, c-MYC, and KLF4 – factors known to induce pluripotency in cells.
  • The team then analyzed the morphology of these eiPSCs, evaluated their potential to maintain pluripotency, and ran a thorough miRNA profile analysis.

Research Findings

  • The team reported successful isolation of multipotent cells. However, they faced failure when trying to generate iPSCs from bone marrow mesenchymal cells.
  • iPSCs derived from adipose tissue, umbilical cord tissue, and adult fibroblasts were found to be positive for alkaline phosphatase, OCT4, and NANOG (markers of pluripotency), and dependent exclusively on bFGF.
  • The generated iPSCs were also capable of forming embryoid bodies – another indication of pluripotency.
  • The profile of miRNAs, small RNA molecules involved in gene regulation, revealed distinct patterns between pluripotent and multipotent cells. Higher levels of certain types of miRNA were found in iPSCs, while other types were prevalent in nonpluripotent (multipotent) cells.

Conclusion

  • The research demonstrated the successful generation of equine iPSCs from adult fibroblasts, adipose, and umbilical cord tissue using human pluripotency factors.
  • The miRNA profile also suggested that different cell lines might respond differently to the reprogramming process.
  • This study provides important insights into equine stem cell biology and paves the way for future research and potential therapeutic applications.

Cite This Article

APA
Pessôa LVF, Pires PRL, Del Collado M, Pieri NCG, Recchia K, Souza AF, Perecin F, da Silveira JC, de Andrade AFC, Ambrosio CE, Bressan FF, Meirelles FV. (2019). Generation and miRNA Characterization of Equine Induced Pluripotent Stem Cells Derived from Fetal and Adult Multipotent Tissues. Stem Cells Int, 2019, 1393791. https://doi.org/10.1155/2019/1393791

Publication

Stem cells international
ISSN: 1687-966X
NlmUniqueID: 101535822
Country: United States
Language: English
Volume: 2019
Pages: 1393791
PII: 1393791

Researcher Affiliations

Pessôa, Laís Vicari de Figueiredo
  • Departamento de Medicina Veterinária, Faculdade de Zootecnia e Engenharia de Alimentos, Universidade de São Paulo, Pirassununga 13635-000, Brazil.
  • Department of Veterinary and Animal Sciences, Section for Anatomy & Biochemistry, University of Copenhagen, 1870 Frederiksberg C, Denmark.
Pires, Pedro Ratto Lisboa
  • Departamento de Medicina Veterinária, Faculdade de Zootecnia e Engenharia de Alimentos, Universidade de São Paulo, Pirassununga 13635-000, Brazil.
Del Collado, Maite
  • Departamento de Medicina Veterinária, Faculdade de Zootecnia e Engenharia de Alimentos, Universidade de São Paulo, Pirassununga 13635-000, Brazil.
Pieri, Naira Caroline Godoy
  • Departamento de Reprodução Animal, Faculdade de Medicina Veterinária e Zootecnia, Universidade de São Paulo, Pirassununga 13635-000, Brazil.
Recchia, Kaiana
  • Departamento de Medicina Veterinária, Faculdade de Zootecnia e Engenharia de Alimentos, Universidade de São Paulo, Pirassununga 13635-000, Brazil.
Souza, Aline Fernanda
  • Departamento de Medicina Veterinária, Faculdade de Zootecnia e Engenharia de Alimentos, Universidade de São Paulo, Pirassununga 13635-000, Brazil.
Perecin, Felipe
  • Departamento de Medicina Veterinária, Faculdade de Zootecnia e Engenharia de Alimentos, Universidade de São Paulo, Pirassununga 13635-000, Brazil.
da Silveira, Juliano Coelho
  • Departamento de Medicina Veterinária, Faculdade de Zootecnia e Engenharia de Alimentos, Universidade de São Paulo, Pirassununga 13635-000, Brazil.
de Andrade, André Furugen Cesar
  • Departamento de Reprodução Animal, Faculdade de Medicina Veterinária e Zootecnia, Universidade de São Paulo, Pirassununga 13635-000, Brazil.
Ambrosio, Carlos Eduardo
  • Departamento de Medicina Veterinária, Faculdade de Zootecnia e Engenharia de Alimentos, Universidade de São Paulo, Pirassununga 13635-000, Brazil.
Bressan, Fabiana Fernandes
  • Departamento de Medicina Veterinária, Faculdade de Zootecnia e Engenharia de Alimentos, Universidade de São Paulo, Pirassununga 13635-000, Brazil.
Meirelles, Flavio Vieira
  • Departamento de Medicina Veterinária, Faculdade de Zootecnia e Engenharia de Alimentos, Universidade de São Paulo, Pirassununga 13635-000, Brazil.

References

This article includes 35 references
  1. Kassem M, Kristiansen M, Abdallah BM. Mesenchymal stem cells: cell biology and potential use in therapy.. Basic Clin Pharmacol Toxicol 2004 Nov;95(5):209-14.
    doi: 10.1111/j.1742-7843.2004.pto950502.xpubmed: 15546474google scholar: lookup
  2. Carrade DD, Lame MW, Kent MS, Clark KC, Walker NJ, Borjesson DL. Comparative Analysis of the Immunomodulatory Properties of Equine Adult-Derived Mesenchymal Stem Cells().. Cell Med 2012;4(1):1-11.
    doi: 10.3727/215517912X647217pmc: PMC3495591pubmed: 23152950google scholar: lookup
  3. Uder C, Brückner S, Winkler S, Tautenhahn HM, Christ B. Mammalian MSC from selected species: Features and applications.. Cytometry A 2018 Jan;93(1):32-49.
    doi: 10.1002/cyto.a.23239pubmed: 28906582google scholar: lookup
  4. Saito S, Ugai H, Sawai K, Yamamoto Y, Minamihashi A, Kurosaka K, Kobayashi Y, Murata T, Obata Y, Yokoyama K. Isolation of embryonic stem-like cells from equine blastocysts and their differentiation in vitro.. FEBS Lett 2002 Nov 20;531(3):389-96.
    doi: 10.1016/S0014-5793(02)03550-0pubmed: 12435581google scholar: lookup
  5. Li X, Zhou SG, Imreh MP, Ahrlund-Richter L, Allen WR. Horse embryonic stem cell lines from the proliferation of inner cell mass cells.. Stem Cells Dev 2006 Aug;15(4):523-31.
    doi: 10.1089/scd.2006.15.523pubmed: 16978056google scholar: lookup
  6. Nagy K, Sung HK, Zhang P, Laflamme S, Vincent P, Agha-Mohammadi S, Woltjen K, Monetti C, Michael IP, Smith LC, Nagy A. Induced pluripotent stem cell lines derived from equine fibroblasts.. Stem Cell Rev Rep 2011 Sep;7(3):693-702.
    doi: 10.1007/s12015-011-9239-5pmc: PMC3137777pubmed: 21347602google scholar: lookup
  7. Khodadadi K, Sumer H, Pashaiasl M, Lim S, Williamson M, Verma PJ. Induction of pluripotency in adult equine fibroblasts without c-MYC.. Stem Cells Int 2012;2012:429160.
    doi: 10.1155/2012/429160pmc: PMC3328202pubmed: 22550508google scholar: lookup
  8. Breton A, Sharma R, Diaz AC, Parham AG, Graham A, Neil C, Whitelaw CB, Milne E, Donadeu FX. Derivation and characterization of induced pluripotent stem cells from equine fibroblasts.. Stem Cells Dev 2013 Feb 15;22(4):611-21.
    doi: 10.1089/scd.2012.0052pmc: PMC3564467pubmed: 22897112google scholar: lookup
  9. Whitworth DJ, Ovchinnikov DA, Sun J, Fortuna PR, Wolvetang EJ. Generation and characterization of leukemia inhibitory factor-dependent equine induced pluripotent stem cells from adult dermal fibroblasts.. Stem Cells Dev 2014 Jul 1;23(13):1515-23.
    doi: 10.1089/scd.2013.0461pmc: PMC4066230pubmed: 24555755google scholar: lookup
  10. Sharma R, Livesey MR, Wyllie DJ, Proudfoot C, Whitelaw CB, Hay DC, Donadeu FX. Generation of functional neurons from feeder-free, keratinocyte-derived equine induced pluripotent stem cells.. Stem Cells Dev 2014 Jul 1;23(13):1524-34.
    doi: 10.1089/scd.2013.0565pubmed: 24548115google scholar: lookup
  11. Lee EM, Kim AY, Lee EJ, Park JK, Park SI, Cho SG, Kim HK, Kim SY, Jeong KS. Generation of Equine-Induced Pluripotent Stem Cells and Analysis of Their Therapeutic Potential for Muscle Injuries.. Cell Transplant 2016 Nov;25(11):2003-2016.
    doi: 10.3727/096368916X691691pubmed: 27226077google scholar: lookup
  12. Kim K, Doi A, Wen B, Ng K, Zhao R, Cahan P, Kim J, Aryee MJ, Ji H, Ehrlich LI, Yabuuchi A, Takeuchi A, Cunniff KC, Hongguang H, McKinney-Freeman S, Naveiras O, Yoon TJ, Irizarry RA, Jung N, Seita J, Hanna J, Murakami P, Jaenisch R, Weissleder R, Orkin SH, Weissman IL, Feinberg AP, Daley GQ. Epigenetic memory in induced pluripotent stem cells.. Nature 2010 Sep 16;467(7313):285-90.
    doi: 10.1038/nature09342pmc: PMC3150836pubmed: 20644535google scholar: lookup
  13. Raab S, Klingenstein M, Liebau S, Linta L. A Comparative View on Human Somatic Cell Sources for iPSC Generation.. Stem Cells Int 2014;2014:768391.
    doi: 10.1155/2014/768391pmc: PMC4241335pubmed: 25431601google scholar: lookup
  14. Hackett CH, Greve L, Novakofski KD, Fortier LA. Comparison of gene-specific DNA methylation patterns in equine induced pluripotent stem cell lines with cells derived from equine adult and fetal tissues.. Stem Cells Dev 2012 Jul 1;21(10):1803-11.
    doi: 10.1089/scd.2011.0055pmc: PMC3376462pubmed: 21988203google scholar: lookup
  15. Zhang W, Zhong L, Wang J, Han J. Distinct MicroRNA Expression Signatures of Porcine Induced Pluripotent Stem Cells under Mouse and Human ESC Culture Conditions.. PLoS One 2016;11(7):e0158655.
    doi: 10.1371/journal.pone.0158655pmc: PMC4934789pubmed: 27384321google scholar: lookup
  16. Wilson KD, Venkatasubrahmanyam S, Jia F, Sun N, Butte AJ, Wu JC. MicroRNA profiling of human-induced pluripotent stem cells.. Stem Cells Dev 2009 Jun;18(5):749-58.
    doi: 10.1089/scd.2008.0247pmc: PMC3135181pubmed: 19284351google scholar: lookup
  17. Sharma A, Wu JC. MicroRNA expression profiling of human-induced pluripotent and embryonic stem cells.. Methods Mol Biol 2013;936:247-56.
    doi: 10.1007/978-1-62703-083-0_19pmc: PMC3638037pubmed: 23007513google scholar: lookup
  18. Maia L, De Vita B, Moraes C N, Destro F C, Landim-Alvarenga F C, Amorim R M. Considerações sobre a obtenção, processamento, caracterização e aplicação terapêutica das células-tronco mesenquimais em medicina equina. Veterinária e Zootecnia 2013;20:359–373.
  19. Carrade DD, Owens SD, Galuppo LD, Vidal MA, Ferraro GL, Librach F, Buerchler S, Friedman MS, Walker NJ, Borjesson DL. Clinicopathologic findings following intra-articular injection of autologous and allogeneic placentally derived equine mesenchymal stem cells in horses.. Cytotherapy 2011 Apr;13(4):419-30.
    doi: 10.3109/14653249.2010.536213pubmed: 21105841google scholar: lookup
  20. De Schauwer C, Goossens K, Piepers S, Hoogewijs MK, Govaere JL, Smits K, Meyer E, Van Soom A, Van de Walle GR. Characterization and profiling of immunomodulatory genes of equine mesenchymal stromal cells from non-invasive sources.. Stem Cell Res Ther 2014 Jan 13;5(1):6.
    doi: 10.1186/scrt395pmc: PMC4055120pubmed: 24418262google scholar: lookup
  21. Barberini DJ, Freitas NP, Magnoni MS, Maia L, Listoni AJ, Heckler MC, Sudano MJ, Golim MA, da Cruz Landim-Alvarenga F, Amorim RM. Equine mesenchymal stem cells from bone marrow, adipose tissue and umbilical cord: immunophenotypic characterization and differentiation potential.. Stem Cell Res Ther 2014 Feb 21;5(1):25.
    doi: 10.1186/scrt414pmc: PMC4055040pubmed: 24559797google scholar: lookup
  22. Gruber HE, Somayaji S, Riley F, Hoelscher GL, Norton HJ, Ingram J, Hanley EN Jr. Human adipose-derived mesenchymal stem cells: serial passaging, doubling time and cell senescence.. Biotech Histochem 2012 May;87(4):303-11.
    doi: 10.3109/10520295.2011.649785pubmed: 22250760google scholar: lookup
  23. Sommer CA, Stadtfeld M, Murphy GJ, Hochedlinger K, Kotton DN, Mostoslavsky G. Induced pluripotent stem cell generation using a single lentiviral stem cell cassette.. Stem Cells 2009 Mar;27(3):543-9.
    doi: 10.1634/stemcells.2008-1075pmc: PMC4848035pubmed: 19096035google scholar: lookup
  24. De Bem THC, da Silveira JC, Sampaio RV, Sangalli JR, Oliveira MLF, Ferreira RM, Silva LA, Perecin F, King WA, Meirelles FV, Ramos ES. Low levels of exosomal-miRNAs in maternal blood are associated with early pregnancy loss in cloned cattle.. Sci Rep 2017 Oct 30;7(1):14319.
    doi: 10.1038/s41598-017-14616-1pmc: PMC5662615pubmed: 29085015google scholar: lookup
  25. Xia J, Sinelnikov IV, Han B, Wishart DS. MetaboAnalyst 3.0--making metabolomics more meaningful.. Nucleic Acids Res 2015 Jul 1;43(W1):W251-7.
    doi: 10.1093/nar/gkv380pmc: PMC4489235pubmed: 25897128google scholar: lookup
  26. De Schauwer C, Meyer E, Van de Walle GR, Van Soom A. Markers of stemness in equine mesenchymal stem cells: a plea for uniformity.. Theriogenology 2011 May;75(8):1431-43.
    doi: 10.1016/j.theriogenology.2010.11.008pubmed: 21196039google scholar: lookup
  27. Olivera R, Moro LN, Jordan R, Luzzani C, Miriuka S, Radrizzani M, Donadeu FX, Vichera G. 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.
    doi: 10.1371/journal.pone.0164049pmc: PMC5061425pubmed: 27732616google scholar: lookup
  28. Jin HJ, Bae YK, Kim M, Kwon SJ, Jeon HB, Choi SJ, Kim SW, Yang YS, Oh W, Chang JW. Comparative analysis of human mesenchymal stem cells from bone marrow, adipose tissue, and umbilical cord blood as sources of cell therapy.. Int J Mol Sci 2013 Sep 3;14(9):17986-8001.
    doi: 10.3390/ijms140917986pmc: PMC3794764pubmed: 24005862google scholar: lookup
  29. Aliborzi G, Vahdati A, Mehrabani D, Hosseini SE, Tamadon A. Isolation, Characterization and Growth Kinetic Comparison of Bone Marrow and Adipose Tissue Mesenchymal Stem Cells of Guinea Pig.. Int J Stem Cells 2016 May 30;9(1):115-23.
    doi: 10.15283/ijsc.2016.9.1.115pmc: PMC4961111pubmed: 27426093google scholar: lookup
  30. Kretlow JD, Jin YQ, Liu W, Zhang WJ, Hong TH, Zhou G, Baggett LS, Mikos AG, Cao Y. Donor age and cell passage affects differentiation potential of murine bone marrow-derived stem cells.. BMC Cell Biol 2008 Oct 28;9:60.
    doi: 10.1186/1471-2121-9-60pmc: PMC2584028pubmed: 18957087google scholar: lookup
  31. Zaim M, Karaman S, Cetin G, Isik S. Donor age and long-term culture affect differentiation and proliferation of human bone marrow mesenchymal stem cells.. Ann Hematol 2012 Aug;91(8):1175-86.
    doi: 10.1007/s00277-012-1438-xpubmed: 22395436google scholar: lookup
  32. Carter-Arnold JL, Neilsen NL, Amelse LL, Odoi A, Dhar MS. In vitro analysis of equine, bone marrow-derived mesenchymal stem cells demonstrates differences within age- and gender-matched horses.. Equine Vet J 2014 Sep;46(5):589-95.
    doi: 10.1111/evj.12142pubmed: 23855680google scholar: lookup
  33. Ruiz S, Panopoulos AD, Herrerías A, Bissig KD, Lutz M, Berggren WT, Verma IM, Izpisua Belmonte JC. A high proliferation rate is required for cell reprogramming and maintenance of human embryonic stem cell identity.. Curr Biol 2011 Jan 11;21(1):45-52.
    doi: 10.1016/j.cub.2010.11.049pmc: PMC3034649pubmed: 21167714google scholar: lookup
  34. Sandmaier SE, Telugu BP. MicroRNA-Mediated Reprogramming of Somatic Cells into Induced Pluripotent Stem Cells.. Methods Mol Biol 2015;1330:29-36.
    doi: 10.1007/978-1-4939-2848-4_3pubmed: 26621586google scholar: lookup
  35. Porciuncula A, Zapata N, Guruceaga E, Agirre X, Barajas M, Prosper F. MicroRNA signatures of iPSCs and endoderm-derived tissues.. Gene Expr Patterns 2013 Jan-Feb;13(1-2):12-20.
    doi: 10.1016/j.gep.2012.08.002pubmed: 22982176google scholar: lookup

Citations

This article has been cited 0 times.
Subscribe to our newsletter
Join 99,357 horse owners like you who receive our email updates on horse health and nutrition.