Stem potential of the new human cell line 4BL

  • V. O. Kushniruk Institute of molecular biology and genetics of NAS of Ukraine, Ukraine, 03143, Kyiv, Akademika Zabolotnoho str., 150
  • V. A. Shablii Institute of Cell Therapy, Ukraine, 03680, Kyiv, Kosmonavta Komarova Ave., 3
  • S. P. Shpylevaya Institute of molecular biology and genetics of NAS of Ukraine, Ukraine, 03143, Kyiv, Akademika Zabolotnoho str., 150
  • T. P. Ruban Institute of molecular biology and genetics of NAS of Ukraine, Ukraine, 03143, Kyiv, Akademika Zabolotnoho str., 150
  • O. O. Piven Institute of molecular biology and genetics of NAS of Ukraine, Ukraine, 03143, Kyiv, Akademika Zabolotnoho str., 150
  • G. S. Lobyntseva Institute of Cell Therapy, Ukraine, 03680, Kyiv, Kosmonavta Komarova Ave., 3
  • L. L. Lukash Institute of molecular biology and genetics of NAS of Ukraine, Ukraine, 03143, Kyiv, Akademika Zabolotnoho str., 150
DOI: https://doi.org/10.7124/visnyk.utgis.14.1.541

Abstract

During generation of new cell line the researchers are primarily interested in such characteristics as morphology and immunophenotype, which allows attributing beloning of the cell to that or other type and accordingly outlining branches of its application. The aim of this research was to test the stem potential of the new cell line 4BL, which was derived from the peripheral blood of healthy donor and to investigate it`s immunophenotype. Methods. The standard cell cultivation and the soft agar test were used. Stem potential was checked by differentiation in the adypogenic, osteogenic and myogenic directions. Results. The 4BL cell lines form colonies similar to embryoid bodies when grown in semisolid agar and capable to differentiate into osteogenic, myogenic and adypohenic direction when grown in induction media. Over 90 % of the 4BL line cell population expressed stem cell markers CD105 and C73 and were negative for the hemopoietic cell markers CD90, CD45, C34 and CD14. Also these cells didn’t express Oct 4. Conclusions. The new human cell line 4BL has stem potential and, most likely, belongs to non-hemopoietic multipotent stem cells.

Keywords: cell line, stem cells, embryoid bodies, differentiation, flow cytometry.

References

Tong Z., Solanki A., Hamilos A. et al. Application of biomaterials to advance induced pluripotent stem cell research and therapy. EMBO J. 2015. Vol. 34(8). P. 987–1008. doi: 10.15252/embj.201490756

Chen A., Ting S., Seow J., Reuveny S., Oh S. Considerations in designing systems for large scale production of human cardiomyocytes from pluripotent stem cells. Stem. Cell Res. Ther. 2014. Vol. 5(1). P. 1–12. doi: 10.1186/scrt401

Ramdasi S., Sarang S., Viswanathan C. Potential of mesenchymal stem cell based application in cancer. Int. J. Hematol. Oncol. Stem Cell Res. 2015. Vol. 9(2). P. 95–103.

Hu C., Li L. Two effective routes for removing lineage restriction roadblocks: from somatic cells to hepatocytes. Int. J. Mol. Sci. 2015. Vol. 16(9). P. 20873–20895. doi: 10.3390/ijms160920873

Périé S., Trollet C., Mouly V. et al. Autologous myoblast transplantation for oculopharyngeal muscular dystrophy: a phase I/IIa clinical study. Mol. Ther. 2014. Vol. 22(1). P. 219–225. doi: 10.1038/mt.2013.155

Feng R., Lengner C. Application of stem cell technology in dental regenerative medicine. Adv. Wound Care (New Rochelle). 2013. Vol. 2(6). P.296–305. doi: 10.1089/wound.2012.0375

Han F., Baremberg D., Gao J. et al. Development of stem cell-based therapy for Parkinson's disease. Transl. Neurodegener. 2015. Vol. 4(16). P. 1–13. doi: 10.1186/s40035-015-0039-8

Rezania A., Bruin J.E., Arora P. et al. Reversal of diabetes with insulin-producing cells derived in vitro from human pluripotent stem cells. Nat. Biotechnol. 2014. Vol. 2(11). P. 1121–1133. doi: 10.1038/nbt.3033

Giri S., Bader A. A low-cost, high-quality new drug discovery process using patient-derived induced pluripotent stem cells. Drug Discov. Today. 2015. Vol. 20(1). P. 37–49. doi: 10.1016/j.drudis.2014.10.011

Khetani S.R., Berger D.R., Ballinger K.R. et al. Microengineered liver tissues for drug testing. World J. Stem Cells. 2015. Vol. 7(2). P. 461–469. doi: 10.1177/2211068214566939

King N.M., Perrin J. Ethical issues in stem cell research and therapy. Stem Cell Res. Ther. 2014. Vol. 5(4). P. 85. doi: 10.1186/scrt474

Lukash L. L., Yatsychina A. P., Kushniruk V. O., Pidpala O. V. Reprogrammirovanie somaticheskikh kletok vzroslogo cheloveka. Fakt. Exp. Evol. Org. 2011. Vol. 11. P. 493–498.

Akopyan H. R., Huleyuk N. L., Kushniruk V. O., Mykytenko D. O., Iatsyshyna A. P., Lukash L. L. Comparative analysis of the karyotype of new human cell line 4BL at long-term cultivation: Ploidy of the chromosomal set. Cytology and Genetics. 2013. Vol. 47(5). P. 305-317. doi: 10.3103/S0095452713050022

Macewicz L.L., Kushniruk V.O., Iatsyshyna A.P. et al. Correlation the level of mutagenesis with expression of reparative enzyme O6-metylhuanin DNA methyltransferase (MGMT) during establishment of cell lines in vitro. Biopolymers and cell. 2013. Vol. 29(6). P. 485 492. doi: 10.7124/bc.00083D

Kushniruk V. O., Ruban T. P., Lukash L. L. Morphological and growth peculiarities of new human cell line 4BL. Fakt. Exp. Evol. Org. 2013. Vol. 13. P. 315–319.

Freshney R.I. Culture of animal cells: a manual of basic technique and specialized applications, 6th ed. New Jersey, USA. 2010. 796 p. doi: 10.1002/9780470649367

Hamburger A.W. The human tumor clonogenic assay as a model system in cell biology. Int. J. Cell Cloning. 1987. Vol. 5(2). P. 89–107. doi: 10.1002/stem.5530050202

Amiri F., Halabian R., Salimian M. et al. Induction of multipotency in umbilical cord-derived mesenchymal stem cells cultivated under suspension conditions. Cell Stress Chaperones. 2014. Vol. 19(5). P. 657–666. doi: 10.1007/s12192-014-0491-x

Atlasi Y., Mowla S.J., Ziaee S.A., Bahrami A.R. OCT-4, an embryonic stem cell marker, is highly expressed in bladder cancer. Int. J. Cancer. 2007. Vol. 120(7). P. 1598–1602. doi: 10.1002/ijc.22508

Wang D., Lu P., Zhang H. et al. Oct-4 and Nanog promote the epithelial-mesenchymal transition of breast cancer stem cells and are associated with poor prognosis in breast cancer patients. Oncotarget. 2014. Vol. 5(21). P. 10803–10815. doi: 10.18632/oncotarget.2506

Ode A., Schoon J., Kurtz A. et al. CD73/5'-ecto-nucleotidase acts as a regulatory factor in osteo-/chondrogenic differentiation of mechanically stimulated mesenchymal stromal cells. Eur. Cell Mater. 2013. Vol. 25. P.37–47. doi: 10.22203/eCM.v025a03

Chatterjee D., Tufa D.M., Baehre H. et al. Natural killer cells acquire CD73 expression upon exposure to mesenchymal stem cells. Blood. 2014. Vol. 123(4). P. 594–595. doi: 10.1182/blood-2013-09-524827

Kays S.K., Kaufmann K.B., Abel T. et al. CD105 is a surface marker for receptor-targeted gene transfer into human long-term repopulating hematopoietic stem cells. Stem Cells Dev. 2015. Vol. 24(6). P. 714–723. doi: 10.1089/scd.2014.0455

Rege T.A., Hagood J.S. Thy-1 as a regulator of cell-cell and cell-matrix interactions in axon regeneration, apoptosis, adhesion, migration, cancer, and fibrosis. FASEB J. 2006. Vol. 20(8). P. 1045–1054. doi: 10.1096/fj.05-5460rev

Boxall S.A., Jones E. Markers for characterization of bone marrow multipotential stromal cells. Stem Cells Int. 2012. ID 975871. doi: 10.1155/2012/975871

Shablii V. A., Kuchma M. D., Kyryk V. M. et al. Mesenchymal and trophoblast immunophenotype of multipotent stromal cells from human placenta. Biopolym. Cell. 2014. Vol. 30(2). P. 118–121. doi: 10.7124/bc.000889

Kwon S.M., Lee J.H., Lee S.H. et al. Cross talk with hematopoietic cells regulates the endothelial progenitor cell differentiation of CD34 positive cells. PLoS One. 2014. Vol. 9(8). e106310. doi: 10.1371/journal.pone.0106310

Cyster J.G., Healy J.I., Kishihara K. et al. Regulation of B-lymphocyte negative and positive selection by tyrosine phosphatase CD45. Nature. 1996. Vol. 381(6580). P. 325–328. doi: 10.1038/381325a0

Tarzi R.M., Liu J., Schneiter S. et al. CD14 expression is increased on monocytes in patients with anti-neutrophil cytoplasm antibody (ANCA)-associated vasculitis and correlates with the expression of ANCA autoantigens. Clin. Exp. Immunol. 2015. Vol. 181(1). P.65–75. doi: 10.1111/cei.12625