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The changes of multipotent mesenchymal stromal cells isolated from human adipose tissue during long-term cultivation

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Abstract

Multipotent mesenchymal stromal cells (MMSCs) isolated from human adipose tissue have been subcultured for over 50 population doublings. In three cultures that experienced more than 50 cellular doublings, there appeared cells lacking contact inhibition upon reaching the monolayer. Karyotype analysis (GTG banding) showed a normal diploid karyotype, and aneuploidy and restructurings were not registered. Flow-cytometric analysis of 20 surface antigens on MMSCs in early and late passages revealed changes in the share of cells positively stained with antibodies against CD10 (a zinc depended metalloproteinase); CD34 (sialomucin); CD49 a, d, f, (α1, α4, α6, integrins); and CD71 (a transferrin receptor). Long-term cultivation influenced cell adhesion to proteins of the extracellular matrix (ECM), such as fibronectin and laminin (ligands to α, integrins), as well as the functional abilities of MMSCs to form cells of adipose and bone tissues in vitro. These findings extend our knowledge of cell behavior in culture and allow us to get closer to a deeper understanding of the processes happening to precursor cells in vitro.

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Abbreviations

MMSC:

multipotent mesenchymal stromal cell

SAT:

subcutaneous adipose tissue

FCS:

fetal calf serum

References

  • Aisen, P., Transferrin receptor 1, Int. J. Biochem. Cell Biol., 2004, vol. 36, pp. 2137–2143.

    Article  CAS  PubMed  Google Scholar 

  • Astori, G., Vignati, F., Bardelli, S., Tubio, M., Gola, M., Albertini, V., Bambi, F., Scali, G., Castelli, D., Rasini, V., Soldati, G., and Moccetti, T., “In Vitro” and multicolor phenotypic characterization of cell subpopulations identified in fresh human adipose tissue stromal vascular fraction and in the derived mesenchymal stem cells, J. Transl. Med., 2007, vol. 5, p. 55. doi 10.1186/1479-5876-5-55

    Article  PubMed  PubMed Central  Google Scholar 

  • Baer, P.C. and Geiger, H., Adipose-derived mesenchymal stromal/stem cells: tissue localization, characterization, and heterogeneity, Stem Cells Int., 2012, p. 812693. doi 10.1155/2012/812693

    Google Scholar 

  • Bajek, A., Gurtowska, N., Gackowska, L., Kubiszewska, I., Bodnar, M., Marsza, A., Januszewski, R., Michalkiewicz, J., and Drewa, T., Does the liposuction method influence the phenotypic characteristic of human adipose-derived stem cells?, Biosci. Rep., 2015, vol. 35, art. e00212. doi 10.1042/BSR20150067

    PubMed  PubMed Central  Google Scholar 

  • Bourin, P., Bunnell, B.A., Casteilla, L., Dominici, M., Katz, A.J., March, K.L., Redl, H., Rubin, U.J.P., Yoshimura, K., and Gimble, J.M., Stromal cells from the adipose tissue-derived stromal vascular fraction and culture expanded adipose tissue-derived stromal/stem cells: a joint statement of the international federation for adipose therapeutics (IFATS) and science and the international society for cellular therapy (ISCT), Cytotherapy, 2013, vol. 15, pp. 641–648.

    Article  PubMed  PubMed Central  Google Scholar 

  • Braham, H., Trimeche, M., Ziadi, S., Mestiri, S., Mokni, M., and Amara, K., CD10 expression by fusiform stromal cells in nasopharyngeal carcinoma correlates with tumor progression, Virchows Arch., 2006, vol. 449, pp. 220–224.

    Article  CAS  PubMed  Google Scholar 

  • Guilak, F., Lott, K.E., Awad, H.A., Cao, Q., Hicok, K.C., Fermor, B., and Gimble, J.M., Clonal analysis of the differentiation potential of human adipose-derived adult stem cells, J. Cell Physiol., 2006, vol. 206, pp. 229–237.

    Article  CAS  PubMed  Google Scholar 

  • Huang, W.B., Zhou, X.J., Chen, J.Y., Zhang, L.H., Meng, K., and Ma, H.H., CD10-positive stromal cells in gastric carcinoma: correlation with invasion and metastasis, Jpn. J. Clin. Oncolю, 2005, vol. 35, pp. 245–250.

    Article  Google Scholar 

  • Jeong, S.G. and Cho, G.W., Accumulation of apoptosisinsensitive human bone marrow-mesenchymal stromal cells after long-term expansion, Cell Biochem. Funct., 2016, vol. 22. doi 10.1002/cbf.3191

    Google Scholar 

  • Kim, J., Kang, J.W., Park, J.H., Choi, Y., Choi, K.S., Park, K.D., Baek, D.H., Seong, S.K., Min, H.K., and Kim, H.S., Biological characterization of long-term cultured human mesenchymal stem cells, Arch. Pharm. Res., 2009, vol. 32, pp. 117–126.

    Article  CAS  PubMed  Google Scholar 

  • Lin, C.S., Xin, Z.C., Deng, C.H., Ning, H., Lin, G., and Lue, T.F., Defining adipose tissue-derived stem cells in tissue and in culture, Histol. Histopathol., 2010, vol. 25, pp. 807–815.

    PubMed  Google Scholar 

  • Masloub, S.M., Abdel-Azim, A.M., and Elhamid, E.S., CD10 and osteopontin expression in dentigerous cyst and ameloblastoma, Diagn. Pathol., 2011, vol. 6, p. 44. doi 10.1186/1746-1596-6-44

    Article  PubMed  PubMed Central  Google Scholar 

  • Maumus, M., Peyrafitte, J.A., D’ Angelo, R., Fournier-Wirth, C., Bouloumié, A., Casteilla, L., Sengenès, C., and Bourin, P., Native human adipose stromal cells: localization, morphology and phenotype, Int. J. Obes., 2011, vol. 35, pp. 1141–1153.

    Article  CAS  Google Scholar 

  • Mitchell, J.B., McIntosh, K., Zvonic, S., Garrett, S., Floyd, Z.E., Kloster, A., Di Halvorsen, Y., Storms, R.W., Goh, B., Kilroy, G., Wu, X., and Gimble, J.M., Immunophenotype of human adipose derived cells: temporal changes in stromal-and stem cell-associated markers, Stem Cells, 2006, vol. 24, pp. 376–385.

    Article  PubMed  Google Scholar 

  • Oba, J., Nakahara, T., Hayashida, S., Kido, M., Xie, L., and Takahara, M., Expression of CD10 predicts tumor progression and unfavorable prognosis in malignant melanoma, J. Am. Acad. Dermatol., 2011, vol. 65, pp. 1152–1160.

    Article  CAS  PubMed  Google Scholar 

  • Ogawa, H., Iwaya, K., Izumi, M., Kuroda, M., Serizawa, H., and Koyanagi, Y., Expression of CD10 by stromal cells during colorectal tumor development, Hum. Pathol., 2002, vol. 33, pp. 806–811.

    Article  PubMed  Google Scholar 

  • Redaelli, S., Bentivegna, A., Foudah, D., Miloso, M., Redondo, J., Riva, G., Baronchelli, S., Dalprà, L., and Tredici, G., From cytogenomic to epigenomic profiles: monitoring the biologic behavior of in vitro cultured human bone marrow mesenchymal stem cells, Stem Cell Res. Ther., 2012, vol. 3, p. 47.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Savchenkova, I.P. and Savchenkova, E.A., Integrin expression during long-term cultivation in multipotent mesenchymal stromal cells isolated from human adipose tissue, Cell Tissue Biol., 2014, vol. 8, no. 6, pp. 454–460.

    Article  Google Scholar 

  • Savchenkova, I.P., Rostovskaya, M.S., Chupikova, N.I., Sharifullina, S.Z., and Teplyashin, A.S., Osteogenic differentiation of multipotent mesenchymal stromal cells isolated from human bone marrow and subcutaneous adipose tissue, Cell Tissue Biol., 2008, vol. 2, no. 6, pp. 566–571.

    Article  Google Scholar 

  • Tepliashin, A.S., Korzhikova, S.V., Sharifullina, S.Z., Chupikova, N.I., Rostovskaia, M.S., and Savchenkova, I.P., Characteristics of human mesenchymal stem cells isolated from bone marrow and adipose tissue, Tsitologiia, 2005a, vol. 47, no. 2, pp. 130–135.

    CAS  PubMed  Google Scholar 

  • Tepliashin, A.S., Chupikova, N.I., Korzhikova, S.V., Sharifullina, S.Z., Rostovskaia, M.S., Topchiashvili, Z.A., and Savchenkova, I.P., Comparative analysis of cell populations with a phenotype similar to that of mesenchymal stem cells derived from subcutaneous fat, Tsitologiia, 2005b, vol. 47, no. 7, pp. 637–643.

    CAS  PubMed  Google Scholar 

  • Teplyashin, A.S., Tchyupikova, N.I., Sharifullina, S.Z., Korzhikova, S.V., Rostovskaya, M.S., Topchiashvilly, Z.A., and Savtchenkova, I.P., CD34+ and CD34-MSC-like cells, derived from adipose tissue of human, in 7th Int. Congress Cell Transplant Society. Boston, USA, November 17–20, 2004, 2004a, p. 76.

    Google Scholar 

  • Teplyashin, A.S., Tchupikova, N.I., Sharifullina, S.Z., Rostovskaya, M.S., Korjikova, S.V., Abdrakhmanov, I.K., Dunaev, A.A., Petrin, S.A., Eremenko, V.S., and Savchenkova, I.P., Characterization of human MSC-like cells isolated from bone marrow, adipose tissue, skin and placenta, Herbsttagung der Gesellschaft fur Biochemie und Molekularbiologie, Munster, 2004b, p. 24.

    Google Scholar 

  • Tonti, G.A. and Mannello, F., From bone marrow to therapeutic applications: different behaviour and genetic/epigenetic stability during mesenchymal stem cell expansion in autologous and foetal bovine sera?, Int. J. Dev. Biol., 2008, vol. 52, pp. 1023–1032.

    Article  PubMed  Google Scholar 

  • Traktuev, D.O., Merfeld-Clauss, S., Li, J., Kolonin, M., Arap, W., Pasqualini, R., Johnstone, B.H., and March, K.L., A population of multipotent CD34-positive adipose stromal cells share pericyte and mesenchymal surface markers, reside in a periendothelial location, and stabilize endothelial networks, Circ. Res., 2008, vol. 102, pp. 77–85.

    Article  CAS  PubMed  Google Scholar 

  • Uzbekov, R.E., Analysis of protein expression level dynamics in the cell cycle using synchronized cells, Biochemistry (Moscow), 2004, vol. 69, no. 5, pp. 485–496.

    Article  CAS  Google Scholar 

  • Vacanti, V., Kong, E., Suzuki, G., Sato, K., Canty, J.M., and Lee, T., Phenotypic changes of adult porcine mesenchymal stem cells induced by prolonged passaging in culture, J. Cell Physiol., 2005, vol. 205, pp. 194–201.

    Article  CAS  PubMed  Google Scholar 

  • Varma, M.J., Breuls, R.G., Jurgens, W.J., Bontkes, H.J., Schuurhuis, G.J., van Ham, S.M., and van Milligen, F.J., Phenotypical and functional characterization of freshly isolated adipose tissue-derived stem cells, Stem Cells Dev., 2007, vol. 16, pp. 91–104.

    Article  PubMed  Google Scholar 

  • Wagner, W., Implications of long-term culture for mesenchymal stem cells: genetic defects or epigenetic regulation?, Stem Cell Res. Ther., 2012, vol. 3, p. 54.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Yoshimura, K., Shigeura, T., Matsumoto, D., Sato, T., Takaki, Y., Aiba-Kojima, E., Sato, K., Inoue, K., Nagase, T., Koshima, I., and Gonda, K., Characterization of freshly isolated and cultured cells derived from the fatty and fluid portions of liposuction aspirates, J. Cell. Physiol., 2006, vol. 208, pp. 64–76.

    Article  CAS  PubMed  Google Scholar 

  • Zaim, M., Karaman, S., Cetin, G., and Isik, S., Donor age and long-term culture affect differentiation and proliferation of human bone marrow mesenchymal stem cells, Ann. Hematol., 2012, vol. 91, pp. 1175–1186.

    Article  PubMed  Google Scholar 

  • Zimmerlin, L., Donnenberg, V.S. Rubin, J.P., and Donnenberg, A.D., Mesenchymal markers on human adipose stem/progenitor cells, Cytometry A, 2013, vol. 83, pp. 134–140.

    Article  PubMed  Google Scholar 

  • Zuk, P.A., Zhu, M., Mizuno, H., Huang, J., Futrell, J.W., Katz, A.J., Benhaim, P., Lorenz, H.P., and Hedrick, M.H., Multilineage cells from human adipose tissue: implications for cell-based therapies, Tiss. Eng., 2001, vol. 7, pp. 211–228.

    Article  CAS  Google Scholar 

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Authors and Affiliations

  1. Kovalenko All-Russian State Research Institute of Experimental Veterinary Medicine, Russian Academy of Sciences, Moscow, 109428, Russia

    I. P. Savchenkova, E. A. Savchenkova & M. I. Gulyukin

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  1. I. P. Savchenkova
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  2. E. A. Savchenkova
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  3. M. I. Gulyukin
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Correspondence to I. P. Savchenkova.

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Original Russian Text © I.P. Savchenkova, E.A. Savchenkova, M.I. Gulyukin, 2017, published in Tsitologiya, 2017, Vol. 59, No. 5, pp. 307–314.

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Savchenkova, I.P., Savchenkova, E.A. & Gulyukin, M.I. The changes of multipotent mesenchymal stromal cells isolated from human adipose tissue during long-term cultivation. Cell Tiss. Biol. 11, 349–355 (2017). https://doi.org/10.1134/S1990519X17050066

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  • DOI: https://doi.org/10.1134/S1990519X17050066

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