THE INFLUENCE OF SURFACES MODIFIED BY DIFFERENT POLYMERIC SUBSTANCES ON THE GROWTH OF MOUSE EMBRYONIC FIBROBLASTS CELL LINE NIH3T3

Bìol. Tvarin, 2017, Volume 19, Issue 2, pp. 121–126

http://doi.org/10.15407/animbiol19.02.121

THE INFLUENCE OF SURFACES MODIFIED BY DIFFERENT POLYMERIC SUBSTANCES ON THE GROWTH OF MOUSE EMBRYONIC FIBROBLASTS CELL LINE NIH3T3

O. Shtapenko

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Institute of Animal Biology NAAS, 38 Stusa str., Lviv 79034, Ukraine

Cultivating cells on synthetic surfaces has ensured the most approximated to in vivo conditions, cell growth, migration, differentiation, synthesis of the extracellular matrix components. Cell proliferation and viability are affected by surface chemical and phase composition, surface conformation, surface morphology and specific chemical groups on the surface of the scaffold. In our study, we investigated the effects of modified nanosurfaces, a grafted by polymeric of dextran, APTES and albumin on the proliferation and metabilitic activity of the cells-fibroblasts of NIH3T3 line.

For experiments, NIH3T3 cell line was cultured in medium RPMI 1640 with 10 % FCS, 1 % penicillin-streptomycin in 5 % CO₂ at 37 °C for 72 hours. Cells were seeded at the glass plates, which modified nanolayers in various combinations: Glass/APTES, Glass/APTES/dextran, Glass/APTES/albumin, Glass/albumin, Glass/APTES/dextran/albumin; control group — clean glass. During the study the intensity of proliferative growth of NIH3T3 cell line and its viability to form a monolayer was determined by Trypan blue staining after every 24 h of incubation. For investigation of the metabolic cell parameters (lactate dehydrogenase (LDH) activity, glucose concentration culture medium was collected after 24, 48 and 72 hours of cultivation.

The result indicate that surfaces modified by albumin, APTES/albumin and APTES/dextran/albumin showed a higher proliferative effect on NIH3T3 cell line compared to the surface “native glass” (control group) at 24 hours of cultivation. Surfaces enclosing APTES and mixed APTES/dextran assume a higher viability and metabolic activity of the culture in terms of the activity of LDH and glucose concentration at 48 hours of cultivation. After 72 hours of cultivation, the intensity of cell proliferative growth was seen only in APTES/dextran surface, while in other research groups cells concentration was at the level of the control group.

Obtained results have shown that the glass surface modified with grafted nanolayers of APTES, dextran and albumin is nontoxic and can be used for cultivation of cells.

Keywords: CELL CULTURE, PROLIFERATIVE ACTIVITY, SURFACE, NANOLAYERS, APTES, DEXTRAN, ALBUMIN, CYTOTOXICITY

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