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Effect of Subcultivation of Human Bone Marrow Mesenchymal Stem on their Capacities for Chondrogenesis, Supporting Hematopoiesis, and Telomea Length

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Effects of subcultivation of human bone marrow mesenchymal stem cells on their capacities for chondrogenesis and supporting hematopoiesis, and telomea length were investigated. Mesenchymal stem cells were isolated from human bone marrow aspirates and subcultivated several times at 37 °C under a 5% CO2 atmosphere employing DMEM medium containing 10% FCS up to the 20th population doubling level (PDL). The ratio of CD45− CD105+ cells among these cells slightly increased as PDL increased. However, there was no marked change in the chondrogenic capacity of these cells, which was confirmed by expression assay of aggrecan mRNA and Safranin O staining after pellet cell cultivation. The change in capacity to support hematopoiesis of cord blood cells was not observed among cells with various PDLs. On the other hand, telomere length markedly decreased as PDL increased at a higher rate than that at which telomere length of primary mesenchymal stem cells decreased as the age of donor increased.

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References

  • M. Akiyama Y. Hoshi S. Sakurai H. Yamada O. Yamada H. Mizoguchi (1998a) ArticleTitleChanges of telomere length in children after hematopoietic stem cell transplantation Bone Marrow Transplant. 21 IssueID2 167–171 Occurrence Handle10.1038/sj.bmt.1701060 Occurrence Handle1:STN:280:DyaK1c7kvFakug%3D%3D

    Article  CAS  Google Scholar 

  • M. Akiyama H. Uchiyama Y. Hoshi S. Yano O. Asai Y. Kuraishi O. Yamada H. Mizoguchi H. Yamada (1998b) ArticleTitleChanges of telomere length after hematopoietic stem cell transplantation Exp. Hematol. 26 IssueID8 359–365

    Google Scholar 

  • R.C. Allsopp H. Vaziri C. Patterson S. Goldstein E.V. Younglai A.B. Futcher C.W. Greider C.B. Harley (1992) ArticleTitleTelomere length predicts replicative capacity of human fibroblasts PNAS USA 89 IssueID21 10114–10118 Occurrence Handle1438199 Occurrence Handle1:CAS:528:DyaK3sXhvF2luw%3D%3D

    PubMed  CAS  Google Scholar 

  • A. Banfi A. Muraglia B. Dozin M. Mastrogiacomo R. Cancedda R. Quarto (2000) ArticleTitleProliferation kinetics and differentiation potential of ex vivo expanded human bone marrow stromal cells: implications for their use in cell therapy Exp. Hematol. 28 707–715 Occurrence Handle10.1016/S0301-472X(00)00160-0 Occurrence Handle10880757 Occurrence Handle1:STN:280:DC%2BD3czpsFeisQ%3D%3D

    Article  PubMed  CAS  Google Scholar 

  • F.P. Barry R.E. Boynton S. Haynesworth J.M. Murphy J. Zaia (1999) ArticleTitleThe monoclonal antibody SH-2, raised against human mesenchymal stem cells, recognizes an epitope on endoglin (CD105) Biochem. Biophys. Res. Commun. 265 134–139 Occurrence Handle10.1006/bbrc.1999.1620 Occurrence Handle10548503 Occurrence Handle1:CAS:528:DyaK1MXntVCnu7g%3D

    Article  PubMed  CAS  Google Scholar 

  • H.E. Broxmeyer (1986) ArticleTitleBiomolecule–cell interactions and the regulation of myelopoiesis Int. J. Cell Cloning 4 378–405 Occurrence Handle2431075 Occurrence Handle1:CAS:528:DyaL2sXjtV2rtA%3D%3D

    PubMed  CAS  Google Scholar 

  • S.P. Bruder N. Jaiswal S.E. Haynesworth (1997) ArticleTitleGrowth kinetics, self-renewal, and the osteogenic potential of purified human mesenchymal stem cells during extensive subcultivation and following cryopreservation J. Cell. Biochem. 64 278–294 Occurrence Handle10.1002/(SICI)1097-4644(199702)64:2<278::AID-JCB11>3.0.CO;2-F Occurrence Handle9027588 Occurrence Handle1:CAS:528:DyaK2sXpt1Kqsg%3D%3D

    Article  PubMed  CAS  Google Scholar 

  • S.P. Bruder A.A. Kurth M. Shea W.C. Hayes N. Jaiswal S. Kadiyala (1998) ArticleTitleBone regeneration by implantation of purifiedculture-expanded human mesenchymal stem cells J. Orthop. Res. 16 155–162 Occurrence Handle10.1002/jor.1100160202 Occurrence Handle9621889 Occurrence Handle1:STN:280:DyaK1c3otlajug%3D%3D

    Article  PubMed  CAS  Google Scholar 

  • A.I. Caplan (1991) ArticleTitleMesenchymal stem cells J. Orthop. Res. 9 641–650 Occurrence Handle10.1002/jor.1100090504 Occurrence Handle1870029 Occurrence Handle1:STN:280:By6A3s3nslY%3D

    Article  PubMed  CAS  Google Scholar 

  • A.I. Caplan D.J. Fink T. Goto A.E. Linton R.G. Young S. Wakitani V.M. Goldberg S.E. Haynesworth (1993) Mesenchymal stem cells and tissue repair D.W. Jackson (Eds) The Anterior Cruciate Ligament: Current and Future Concepts Raven Press, Ltd. New York 405–417

    Google Scholar 

  • C.M. Counter A.A. Avilion C.E. LeFeuvre N.G. Stewart C.W. Greider C.B. Harley S. Bacchetti (1992) ArticleTitleTelomere shortening associated with chromosome instability is arrested in immortal cells which express telomerase activity EMBO J. 11 1921–1929 Occurrence Handle1582420 Occurrence Handle1:CAS:528:DyaK38XktVait7g%3D

    PubMed  CAS  Google Scholar 

  • J.E. Dennis A.I. Caplan (1996) ArticleTitleDifferentiation potential of conditionally immortalized mesenchymal progenitor cells from adult marrow of a H-2Kb-tsA58 transgenic mouse J. Cell. Physiol. 167 523–538 Occurrence Handle10.1002/(SICI)1097-4652(199606)167:3<523::AID-JCP16>3.0.CO;2-4 Occurrence Handle8655606 Occurrence Handle1:CAS:528:DyaK28XjtFCls7w%3D

    Article  PubMed  CAS  Google Scholar 

  • T.M. Dexter E. Spooncer (1987) ArticleTitleGrowth and differentiation in the hemopoietic system Annu. Rev. Cell Biol. 3 423–441 Occurrence Handle10.1146/annurev.cb.03.110187.002231 Occurrence Handle3318881 Occurrence Handle1:CAS:528:DyaL1cXkvFemtw%3D%3D

    Article  PubMed  CAS  Google Scholar 

  • K. Dorshkind (1990) ArticleTitleRegulation of hemopoiesis by bone marrow stromal cells and their products Annu. Rev. Immunol. 8 111–137 Occurrence Handle2188660 Occurrence Handle1:CAS:528:DyaK3cXkslGqsL0%3D Occurrence Handle10.1146/annurev.iy.08.040190.000551

    Article  PubMed  CAS  Google Scholar 

  • C.B. Harley A.B. Futcher C.W. Greider (1990) ArticleTitleTelomeres shorten during ageing of human fibroblasts Nature 345 458–460 Occurrence Handle10.1038/345458a0 Occurrence Handle2342578 Occurrence Handle1:CAS:528:DyaK3cXksFKntb4%3D

    Article  PubMed  CAS  Google Scholar 

  • T. Kohler R. Plettig W. Wetzstein B. Schaffer R. Ordemann H.O. Nagels G. Ehninger M. Bornhauser (1999) ArticleTitleDefining optimum conditions for the ex vivo expansion of human umbilical cord blood cells. Influences of progenitor enrichmentinterference with feeder layers, early-acting cytokines and agitation of culture vessels Stem cells 17 IssueID1 19–24 Occurrence Handle10215397 Occurrence Handle1:STN:280:DyaK1M7ktVaktA%3D%3D

    PubMed  CAS  Google Scholar 

  • J. Lustiga S. Kurtz D. Shore (1990) ArticleTitleInvolvement of the silencer and UAS binding-protein RAPI in regulation of telomere length Science 250 IssueID4980 549–553

    Google Scholar 

  • M.K. Majumdar M.A. Thiede J.D. Mosca M. Moorman S.L. Gerson (1998) ArticleTitlePhenotypic and functional comparison of cultures of marrow-derived mesenchymal stem cells (MSCs) and stromal cells J. Cell. Physiol. 176 57–66 Occurrence Handle10.1002/(SICI)1097-4652(199807)176:1<57::AID-JCP7>3.0.CO;2-7 Occurrence Handle9618145 Occurrence Handle1:CAS:528:DyaK1cXjsFGkt7c%3D

    Article  PubMed  CAS  Google Scholar 

  • M.K. Majumdar M.A. Thiede S.E. Haynesworth S.P. Bruder S.L. Gerson (2000) ArticleTitleHuman marrow-derived mesenchymal stem cells (MSCs) express hematopoietic cytokines and support long-term hematopoiesis when differentiated toward stromal and osteogenic lineage J. Hematother. Stem Cell Res. 9 841–848 Occurrence Handle10.1089/152581600750062264 Occurrence Handle11177595 Occurrence Handle1:CAS:528:DC%2BD3MXhtFKgtbg%3D

    Article  PubMed  CAS  Google Scholar 

  • M.F. Pittenger A.M. Mackay S.C. Beck R.K. Jaiswal R. Douglas J.D. Mosca M.A. Moorman D.W. Simonetti S. Craig D.R. Marshak (1999) ArticleTitleMultilineage potential of adult human mesenchymal stem cells Science 28 143–147

    Google Scholar 

  • K.L. Rudolph S. Chang H.W. Lee M. Blasco G.J. Gottlieb C. Greider R.A. DePinho (1999) ArticleTitleLongevity, stress responseand cancer in aging telomerase-deficient mice Cell 96 701–712 Occurrence Handle10.1016/S0092-8674(00)80580-2 Occurrence Handle10089885 Occurrence Handle1:CAS:528:DyaK1MXitVChs7w%3D

    Article  PubMed  CAS  Google Scholar 

  • M. Takagi T. Nakamura C. Matsuda T. Hattori S. Wakitani T. Yoshida (2003) ArticleTitle In vitro proliferation of human bone marrow mesenchymal stem cells employing donor serum and basic fibroblast growth factor Cytotechnology 43 89–96 Occurrence Handle10.1023/B:CYTO.0000039911.46200.61 Occurrence Handle19003212

    Article  PubMed  Google Scholar 

  • K. Takubo N. Izumiyama-Shimomura N. Honma M. Sawabe T. Arai M. Kato M. Oshimura K.I. Nakamura (2002) ArticleTitleTelomere lengths are characteristic in each human individual Exp. Gerontol. 37 IssueID4 523–531 Occurrence Handle10.1016/S0531-5565(01)00218-2 Occurrence Handle11830355 Occurrence Handle1:CAS:528:DC%2BD38XhtVKktLo%3D

    Article  PubMed  CAS  Google Scholar 

  • T. Tsuji Y. Nishimura-Morita Y. Watanabe (1999) ArticleTitleA murine stromal cell line promotes the expansion of CD34high+-primitive progenitor cells isolated from human umbilical cord blood in combination with human cytokines Growth Factors 16 225–240 Occurrence Handle10372963 Occurrence Handle1:STN:280:DyaK1MzgtVWkug%3D%3D

    PubMed  CAS  Google Scholar 

  • C. Verfaillie R. Hurley R. Bhatia (1994) ArticleTitleRole of bone marrow matrix in normal and abnormal hematopoiesis Crit. Rev. Oncol. Hematol. 16 201–224 Occurrence Handle8074802 Occurrence Handle1:STN:280:ByuA2MvpsFU%3D

    PubMed  CAS  Google Scholar 

  • S. Wakitani T. Goto S.J. Pineda R.G. Young J.M. Mansour A.I. Caplan V.M. Goldberg (1994) ArticleTitleMesenchymal cell-based repair of large full-thickness defects of articular-cartilage J. Bone Joint Surg. Am. 76 579–592 Occurrence Handle8150826 Occurrence Handle1:STN:280:ByuB3c7itF0%3D

    PubMed  CAS  Google Scholar 

  • S. Wakitani T. Saito A.I. Caplan (1995) ArticleTitleMyogenic cells derived from rat bone marrow mesenchymal stem cells exposed to 5-azacytidine Muscle Nerve 18 1417–1426 Occurrence Handle10.1002/mus.880181212 Occurrence Handle7477065 Occurrence Handle1:STN:280:BymD2svkvVI%3D

    Article  PubMed  CAS  Google Scholar 

  • M. Yamaguchi F. Hirayama M. Kanai N. Sato K. Fukazawa K. Yamashita K. Sawada K. Koike M. Kuwabara H. Ikeda K. Ikebuchi (2001) ArticleTitleSerum-free coculture system for ex vivo expansion of human cord blood primitive progenitors and SCID mouse-reconstituting cells using human bone marrow primary stromal cells Exp. Hematol. 29 174–182 Occurrence Handle10.1016/S0301-472X(00)00653-6 Occurrence Handle11166456 Occurrence Handle1:CAS:528:DC%2BD3MXptFSrsQ%3D%3D

    Article  PubMed  CAS  Google Scholar 

  • R.G. Young D.L. Butler W. Weber A.I. Caplan S.L. Gordon D.J. Fink (1998) ArticleTitleUse of mesenchymal stem cells in a collagen matrix for Achilles tendon repair J. Orthop. Res. 16 406–413 Occurrence Handle10.1002/jor.1100160403 Occurrence Handle9747780 Occurrence Handle1:STN:280:DyaK1cvhvVGmsA%3D%3D

    Article  PubMed  CAS  Google Scholar 

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  1. Mutsumi Takagi

    Present address: Division of Biotechnology and Molecular Chemistry, Graduate School of Engineering, Hokkaido University, Sapporo, 060-8628, Japan

Authors and Affiliations

  1. International Center for Biotechnology, Osaka University, 2-1 Yamada-oka, Suita, Osaka, 565-0871, Japan

    Masaki Nakahara, Mutsumi Takagi & Toshiomi Yoshida

  2. Department of Orthopedic Surgery, Osaka-Minami National Hospital, 2-1 Kidohigashi-machi, Kawachinagano, Osaka, 586-8521, Japan

    Takako Hattori

  3. Orthopedic Surgery, Shinshu University School of Medicine, 3-1-1 Asahi, Matsumoto, Nagano, 390-8621, Japan

    Shigeyuki Wakitani

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  1. Masaki Nakahara
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Correspondence to Mutsumi Takagi.

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Nakahara, M., Takagi, M., Hattori, T. et al. Effect of Subcultivation of Human Bone Marrow Mesenchymal Stem on their Capacities for Chondrogenesis, Supporting Hematopoiesis, and Telomea Length . Cytotechnology 47, 19–27 (2005). https://doi.org/10.1007/s10616-005-3752-9

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