Potentiation of directed osteogenic differentiation of thymic multipotent stromal cells by prior co-cultivation with thymocytes

Home/2016, Vol. 4, No. 2/Potentiation of directed osteogenic differentiation of thymic multipotent stromal cells by prior co-cultivation with thymocytes

Cell and Organ Transplantology. 2016; 4(2):220-223.
DOI: 10.22494/COT.V4I2.59

Potentiation of directed osteogenic differentiation of thymic multipotent stromal cells by prior co-cultivation with thymocytes

Nikolskiy I. S., Nikolskaya V. V., Demchenko D. L., Zubov D. O.
State Institute of Genetic and Regenerative Medicine NAMS of Ukraine, Kyiv, Ukraine

It is known that multipotent stromal cells (MSCs) and thymocytes possess membrane affinity and interaction in the thymic niches that is essentially important for thymocytes differentiation. However there are no data about possible influence of intercellular contacts in the reverse direction: from the thymocytes to the MSCs.
Materials and methods. The MSCs were obtained from the thymuses of С57ВL mice, using the explants technique, and cultivated under standard conditions during 8-12 passages. Thymocytes or bone marrow cells (106) were added to 4×104 MSCs for 24 hours. Thereafter they were eliminated and standard culture medium was changed by osteogenic or adipogenic differentiation medium and cultured during 10 days. After fixation the cells were stained by 1 % alizarin red S solution or 0.2 % solution of oil red О respectively. After extraction of the stains with 10 % acetic acid or isopropyl alcohol the optic density of extracts at 520 nm was measured.
Results. We found that thymic multipotent stromal cells of the C57BL mice were effectively differentiated in vitro into the osteogenic and adipogenic lineages in the appropriate differentiation media that was evidenced by alizarin red and oil red staining of cell cultures. According to the results of measurement of optic density of the dye extracts, it was found that effectiveness of thymic MSCs differentiation into the osteogenic lineage after prior short-term co-cultivation with the thymocytes is increased.
Conclusions. The contact of thymic stromal cells with thymocytes but not with bone marrow cells in the previous 24 hours potentiates the osteogenic differentiation and has no effect on the adipogenic cells maturation.

Key words: thymic multipotent stromal cells; thymocytes, bone marrow cells; osteogenic differentiation; adipogenic differentiation

Full Text PDF (eng) Full Text PDF (ua)

1. Schofield R. The relationship between the spleen colony-forming cell and the haemopoietic stem cell. Blood Cells. 1978; 4(1-2):7-25.
2. Arai F, Hirao A, Ohmura M, et al. Tie2.Angiopoietin-1 signaling regulates hematopoietic stem cell quiescence in the bone marrow niche. Cell. 2004; 118:149–161.
3. Banwell CM, Partington KM, Jenkinson EJ, et al. Studies on the role of IL-7 presentation by mesenchymal fibroblasts during early thymocyte development. Eur J Immunol. 2000; 30(8):2125-29.
4. Suniara RK, Jenkinson EJ, Owen JJ. An essential role for thymic mesenchyme in early T Cell development. J Exp Med. 2000; 191(6):1051-56.
PMid:10727466 PMCid:PMC2193125
5. Nikolskiy IS, Nikolskaya VV, Savinova VO. Vliyanie vnutrivennogo vvedeniya mul’tipotentnykh stromal’nykh kletok timusa na immunnyy otvet [Intravenous injection of multipotent stromal cells of thymus and immune response]. Vestnik Ural’skoy meditsinskoy akademicheskoy nauki – Journal of Ural Medical Academic Science. 2012; 4:55-56.
6. Barda-Saad M, Rozenszajn LA, Globerson A, et al. Selective adhesion of immature thymocytes to bone marrow stromal cells: relevance to T cell lymphopoiesis. Exp Hematol. 1996; 24(2):386-91.
7. Lotfinegar P, Shamsasenjian K, Movassaghpour A, et al. Immunomodulatory nature and site specific affinity of mesenchymal stem cells: a hope in cell therapy. Adv Pharm Bull. 2014; 4(1):5-13.
8. Li W, Huang G, Su J, et al. Mesenchymal stem cells: a double-endged sword in regulating immune responses. Cell Death Differ. 2012; 19(9):1505-13.
PMid:22421969 PMCid:PMC3422473
9. Le Blank K, Mougiakakos D. Multipotent mesenchymal stromal cells and the innate immune system. Nat Rev Immunol. 2012; 12(5):383-96.
10. Birari F, Lisi V, Pasini A, et al. Immune modulation by mesenchymal stem cells. Med Hemother. 2008; 35(3):194-204.
PMid:21547117 PMCid:PMC3083287
11. Jung Y, Song J, Shiozawa Y, et al. .Hematopoietic stem cells regulate mesenchymal stromal cell induction into osteoblasts thereby participating in the formation of the stem cell niche. Stem Cells. 2008; 26(8):2042-51.
PMid:18499897 PMCid:PMC3513687
12. Osada M, Singh VJ, Wu K, et al. Label retention identifies a multipotent mesenchymal stem cell-like population in the postnatal thymus. PLoS One. 2013; 8(12):e83024. doi: 10.1371.journal.pone.0083024. eCollection 2013
13. Prockop DJ, Phinney DG, Bunnell BA. Mesenchymal stem cells: methods and protocols . Totowa, NJ: Humana Press, 2008. 192 p.

Nikolskiy IS, Nikolskaya VV, Demchenko DL, Zubov DO. Potentiation of directed osteogenic differentiation of thymic multipotent stromal cells by prior co-cultivation with thymocytes. Cell and Organ Transplantology. 2016; 4(2):220-223. doi:10.22494/cot.v4i2.59


Creative Commons License
Is licensed under a Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International License.