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FEBS letters2002; 531(3); 389-396; doi: 10.1016/s0014-5793(02)03550-0

Isolation of embryonic stem-like cells from equine blastocysts and their differentiation in vitro.

Abstract: Embryonic stem (ES) cells are pluripotent cells with the potential capacity to generate any type of cell. We describe here the isolation of pluripotent ES-like cells from equine blastocysts that have been frozen and thawed. Our two lines of ES-like cells (E-1 and E-2) appear to maintain a normal diploid karyotype indefinitely in culture in vitro and to express markers that are characteristic of ES cells from mice, namely, alkaline phosphatase, stage-specific embryonic antigen-1, STAT-3 and Oct 4. After culture of equine ES-like cells in vitro for more than 17 passages, some ES-like cells differentiated to neural precursor cells in the presence of basic fibroblast growth factor (bFGF), epidermal growth factor and platelet-derived growth factor. We also developed a protocol that resulted in the differentiation of ES-like cells in vitro to hematopoietic and endothelial cell lineages in response to bFGF, stem cell factor and oncostatin M. Our observations set the stage for future developments that may allow the use of equine ES-like cells for the treatment of neurological and hematopoietic disorders.
Publication Date: 2002-11-19 PubMed ID: 12435581DOI: 10.1016/s0014-5793(02)03550-0Google Scholar: Lookup
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  • Journal Article
  • Research Support
  • Non-U.S. Gov't

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 describes how stem cells that bear a resemblance to embryonic stem cells can be isolated from horse blastocysts, even after they have been frozen and thawed. These cells can replicate in a lab setting and can differentiate into different types of cells, particularly neural precursor cells and certain types of blood and endothelial cells under specific conditions. This could potentially lead to new approaches for treating neurological and hematopoietic disorders in the future.

Isolation and Maintenance of Embryonic Stem-Like Cells

In the study, the researchers successfully isolated pluripotent embryonic stem-like cells from equine blastocysts, which are early stage embryos. Interestingly, these blastocysts had been previously frozen and thawed. This implies that the blastocysts still retained the vital characteristics that allowed for the harvesting of the pluripotent cells. Key highlights from this section include:

  • The isolated cells were labeled as E-1 and E-2 and demonstrated an ability to sustain a standard diploid karyotype, which refers to a normal chromosomal state, even over an indefinite culture period in a lab setting.
  • These cells also expressed markers characteristic of typical mouse embryonic stem cells such as alkaline phosphatase, stage-specific embryonic antigen-1 (SSEA-1), STAT-3 and Oct 4. These are signifiers of their pluripotent status.

Differentiation of Stem Cells

The differentiation of cells refers to the process in which a cell changes from one type to another. In this context, the researchers observed how the stem cells changed or ‘differentiated’ into other cell types under certain conditions.

  • After culturing the ES-like cells for more than 17 passages, some cells differentiated into neural precursor cells when exposed to basic fibroblast growth factor (bFGF), epidermal growth factor, and platelet-derived growth factor. Neural precursor cells are early-stage cells that have the potential to develop into neurons or other neural cells.
  • A protocol was developed to manipulate the stem cells to differentiate into hematopoietic and endothelial cell lineages. This was done in response to bFGF, stem cell factor and oncostatin M.

Implications for Future Treatment of Disorders

This research establishes a fundamental understanding of how equine stem-like cells can be manipulated in vitro, which could pave the way for future developments in the treatment of neurological and hematopoietic (blood-related) disorders.

  • Development of new therapies or treatments using these stem-like cells could offer hope for conditions that are currently difficult to treat or incurable due to the limitations of the body’s ability to regenerate appropriate cells.
  • Specifically, in horses, this might lead to new treatments for equine neurological disorders or conditions related to their blood circulatory system.

Cite This Article

APA
Saito S, Ugai H, Sawai K, Yamamoto Y, Minamihashi A, Kurosaka K, Kobayashi Y, Murata T, Obata Y, Yokoyama K. (2002). Isolation of embryonic stem-like cells from equine blastocysts and their differentiation in vitro. FEBS Lett, 531(3), 389-396. https://doi.org/10.1016/s0014-5793(02)03550-0

Publication

ISSN: 0014-5793
NlmUniqueID: 0155157
Country: England
Language: English
Volume: 531
Issue: 3
Pages: 389-396

Researcher Affiliations

Saito, Shigeo
  • Gene Engineering Division, BioResource Center, RIKEN, Tsukuba, 305-0074, Ibaraki, Japan.
Ugai, Hideyo
    Sawai, Ken
      Yamamoto, Yusuke
        Minamihashi, Akira
          Kurosaka, Kahori
            Kobayashi, Yoshiro
              Murata, Takehide
                Obata, Yuichi
                  Yokoyama, Kazunari

                    MeSH Terms

                    • Animals
                    • Base Sequence
                    • Biomarkers
                    • Cell Differentiation
                    • Cell Lineage
                    • DNA Primers
                    • Embryo, Mammalian / cytology
                    • Horses
                    • In Vitro Techniques
                    • Mice
                    • Molecular Sequence Data
                    • Reverse Transcriptase Polymerase Chain Reaction
                    • Stem Cells / cytology

                    Citations

                    This article has been cited 31 times.
                    1. Popova J, Bets V, Kozhevnikova E. Perspectives in Genome-Editing Techniques for Livestock.. Animals (Basel) 2023 Aug 10;13(16).
                      doi: 10.3390/ani13162580pubmed: 37627370google scholar: lookup
                    2. Li ZJ, Yang QQ, Zhou YL. Basic Research on Tendon Repair: Strategies, Evaluation, and Development.. Front Med (Lausanne) 2021;8:664909.
                      doi: 10.3389/fmed.2021.664909pubmed: 34395467google scholar: lookup
                    3. Hisey E, Ross PJ, Meyers SA. A Review of OCT4 Functions and Applications to Equine Embryos.. J Equine Vet Sci 2021 Mar;98:103364.
                      doi: 10.1016/j.jevs.2020.103364pubmed: 33663726google scholar: lookup
                    4. Kumar D, Talluri TR, Selokar NL, Hyder I, Kues WA. Perspectives of pluripotent stem cells in livestock.. World J Stem Cells 2021 Jan 26;13(1):1-29.
                      doi: 10.4252/wjsc.v13.i1.1pubmed: 33584977google scholar: lookup
                    5. Shojaee A, Parham A. Strategies of tenogenic differentiation of equine stem cells for tendon repair: current status and challenges.. Stem Cell Res Ther 2019 Jun 18;10(1):181.
                      doi: 10.1186/s13287-019-1291-0pubmed: 31215490google scholar: lookup
                    6. 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. Generation and miRNA Characterization of Equine Induced Pluripotent Stem Cells Derived from Fetal and Adult Multipotent Tissues.. Stem Cells Int 2019;2019:1393791.
                      doi: 10.1155/2019/1393791pubmed: 31191664google scholar: lookup
                    7. 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
                    8. Kim D, Jung YG, Roh S. Microarray analysis of embryo-derived bovine pluripotent cells: The vulnerable state of bovine embryonic stem cells.. PLoS One 2017;12(3):e0173278.
                      doi: 10.1371/journal.pone.0173278pubmed: 28257460google scholar: lookup
                    9. Li S, Lan H, Men H, Wu Y, Li N, Capecchi MR, Bryda EC, Wu S. Derivation of Transgene-Free Rat Induced Pluripotent Stem Cells Approximating the Quality of Embryonic Stem Cells.. Stem Cells Transl Med 2017 Feb;6(2):340-351.
                      doi: 10.5966/sctm.2015-0390pubmed: 28191784google scholar: lookup
                    10. Saito S, Lin YC, Murayama Y, Nakamura Y, Eckner R, Niemann H, Yokoyama KK. Retracted article: In vitro derivation of mammalian germ cells from stem cells and their potential therapeutic application.. Cell Mol Life Sci 2015 Dec;72(23):4545-60.
                      doi: 10.1007/s00018-015-2020-1pubmed: 26439925google scholar: lookup
                    11. Shah SM, Saini N, Ashraf S, Zandi M, Manik RS, Singla SK, Palta P, Chauhan MS. Development, Characterization, and Pluripotency Analysis of Buffalo (Bubalus bubalis) Embryonic Stem Cell Lines Derived from In Vitro-Fertilized, Hand-Guided Cloned, and Parthenogenetic Embryos.. Cell Reprogram 2015 Aug;17(4):306-22.
                      doi: 10.1089/cell.2014.0098pubmed: 26168169google scholar: lookup
                    12. Petkova R, Arabadjiev B, Chakarov S, Pankov R. Current state of the opportunities for derivation of germ-like cells from pluripotent stem cells: are you a man, or a mouse?. Biotechnol Biotechnol Equip 2014 Mar 4;28(2):184-191.
                      doi: 10.1080/13102818.2014.907037pubmed: 26019504google scholar: lookup
                    13. Han JY, Lee HC, Park TS. Germline-competent stem cell in avian species and its application.. Asian J Androl 2015 May-Jun;17(3):421-6.
                      doi: 10.4103/1008-682X.148073pubmed: 25677135google scholar: lookup
                    14. 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.2013.0461pubmed: 24555755google scholar: lookup
                    15. Wang SW, Wang SS, Wu DC, Lin YC, Ku CC, Wu CC, Chai CY, Lee JN, Tsai EM, Lin CL, Yang RC, Ko YC, Yu HS, Huo C, Chuu CP, Murayama Y, Nakamura Y, Hashimoto S, Matsushima K, Jin C, Eckner R, Lin CS, Saito S, Yokoyama KK. Androgen receptor-mediated apoptosis in bovine testicular induced pluripotent stem cells in response to phthalate esters.. Cell Death Dis 2013 Nov 7;4(11):e907.
                      doi: 10.1038/cddis.2013.420pubmed: 24201806google scholar: lookup
                    16. Kumar R, Ahlawat SP, Sharma M, Verma OP, Sai Kumar G, Taru Sharma G. Selection of appropriate isolation method based on morphology of blastocyst for efficient derivation of buffalo embryonic stem cells.. Cytotechnology 2014 Mar;66(2):239-50.
                      doi: 10.1007/s10616-013-9561-7pubmed: 23553019google scholar: lookup
                    17. Singh KP, Kaushik R, Garg V, Sharma R, George A, Singh MK, Manik RS, Palta P, Singla SK, Chauhan MS. Expression pattern of pluripotent markers in different embryonic developmental stages of buffalo (Bubalus bubalis) embryos and putative embryonic stem cells generated by parthenogenetic activation.. Cell Reprogram 2012 Dec;14(6):530-8.
                      doi: 10.1089/cell.2012.0032pubmed: 23194456google scholar: lookup
                    18. 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.2012.0052pubmed: 22897112google scholar: lookup
                    19. Verma OP, Kumar R, Nath A, Sharma M, Dubey PK, Kumar GS, Sharma GT. In vivo differentiation potential of buffalo (Bubalus bubalis) embryonic stem cell.. In Vitro Cell Dev Biol Anim 2012 Jun;48(6):349-58.
                      doi: 10.1007/s11626-012-9515-ypubmed: 22678753google scholar: lookup
                    20. Muzaffar M, Selokar NL, Singh KP, Zandi M, Singh MK, Shah RA, Chauhan MS, Singla SK, Palta P, Manik R. Equivalency of buffalo (Bubalus bubalis) embryonic stem cells derived from fertilized, parthenogenetic, and hand-made cloned embryos.. Cell Reprogram 2012 Jun;14(3):267-79.
                      doi: 10.1089/cell.2011.0090pubmed: 22582863google scholar: lookup
                    21. 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: 10.1155/2012/429160pubmed: 22550508google scholar: lookup
                    22. Hackett CH, Fortier LA. Embryonic stem cells and iPS cells: sources and characteristics.. Vet Clin North Am Equine Pract 2011 Aug;27(2):233-42.
                      doi: 10.1016/j.cveq.2011.04.003pubmed: 21872756google scholar: lookup
                    23. 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/s12015-011-9239-5pubmed: 21347602google scholar: lookup
                    24. Telugu BP, Ezashi T, Roberts RM. The promise of stem cell research in pigs and other ungulate species.. Stem Cell Rev Rep 2010 Mar;6(1):31-41.
                      doi: 10.1007/s12015-009-9101-1pubmed: 19949895google scholar: lookup
                    25. Koch TG, Berg LC, Betts DH. Concepts for the clinical use of stem cells in equine medicine.. Can Vet J 2008 Oct;49(10):1009-17.
                      pubmed: 19119371
                    26. Talbot NC, Blomberg Le Ann. The pursuit of ES cell lines of domesticated ungulates.. Stem Cell Rev 2008 Sep;4(3):235-54.
                      doi: 10.1007/s12015-008-9026-0pubmed: 18612851google scholar: lookup
                    27. Troyer DL, Weiss ML. Wharton's jelly-derived cells are a primitive stromal cell population.. Stem Cells 2008 Mar;26(3):591-9.
                      doi: 10.1634/stemcells.2007-0439pubmed: 18065397google scholar: lookup
                    28. Weiss ML, Troyer DL. Stem cells in the umbilical cord.. Stem Cell Rev 2006;2(2):155-62.
                      doi: 10.1007/s12015-006-0022-ypubmed: 17237554google scholar: lookup
                    29. Saito S, Yokoyama K, Tamagawa T, Ishiwata I. Derivation and induction of the differentiation of animal ES cells as well as human pluripotent stem cells derived from fetal membrane.. Hum Cell 2005 Sep;18(3):135-41.
                    30. Mikkola M, Olsson C, Palgi J, Ustinov J, Palomaki T, Horelli-Kuitunen N, Knuutila S, Lundin K, Otonkoski T, Tuuri T. Distinct differentiation characteristics of individual human embryonic stem cell lines.. BMC Dev Biol 2006 Aug 8;6:40.
                      doi: 10.1186/1471-213X-6-40pubmed: 16895598google scholar: lookup
                    31. Saito S, Liu B, Yokoyama K. Animal embryonic stem (ES) cells: self-renewal, pluripotency, transgenesis and nuclear transfer.. Hum Cell 2004 Sep;17(3):107-15.