Original scientific paper

Organospecific influence of the extract of cryopreserved piglets’ skin fragments

IRYNA BESPALOVA ; Institute for Problems of Cryobiology and Cryomedicine of the National Academy of Sciences of Ukraine, 23 Pereyaslavskaya str., Kharkov, Ukraine 61015
IRYNA BELOCHKINA ; Institute for Problems of Cryobiology and Cryomedicine of the National Academy of Sciences of Ukraine, 23 Pereyaslavskaya str., Kharkov, Ukraine 61015
SERGIY GALCHENKO ; Institute for Problems of Cryobiology and Cryomedicine of the National Academy of Sciences of Ukraine, 23 Pereyaslavskaya str., Kharkov, Ukraine 61015
BORIS SANDOMIRSKY

Abstract

Researching mechanisms of peptides’ tissue-specific effect is an important task of modern molecular biology, physiology and medicine. In previous studies it was shown that extracts of cryopreserved piglets’ skin fragments and pigs’ spleen fragments accelerate and normalise the healing of skin wound in experiment. Fibroblasts culture is an appropriate model for studying tissue-specific biological activity of peptide complexes on relevant type of cells. This research was aimed to establish the influence of the extract of cryopreserved newborn piglets’ skin fragments (cNPSE) and the extract of cryopreserved pigs’ spleen fragments (cPSE) on proliferative and metabolic activity of skin fibroblasts in culture.

Extracts were obtained from cryopreserved skin fragments and cryopreserved spleen fragments. Primary culture of neonatal rat skin fibroblasts was obtained by free cell transfer from skin fragments and subsequent reseeding. Metabolic activity of cells in culture was defined using non-toxic redox indicator AlamarBlue. The number of cells in a well was measured by counting the quantity of cells in wells.


On the 7th day the metabolic activity of fibroblasts, cultured with cNPSE
(1 μg/ml final peptide concentration), was higher than control by 1.3 times. Adding of cPSE (in the same concentration of peptides) increased cell metabolic activity by 1.2 times. While incubating fibroblasts in the medium with 2% FBS, a decrease of metabolic activity of cells was observed on the 5th day, and by the 7th day it was 51.2% of the control. At 1 and 1.5 μg/ml final peptide concentration of cNPSE, the metabolic activity of fibroblasts remained at the level observed in the control samples with 10% FBS. Adding cPSE to the incubation medium did not affect the metabolic activity of cells. Increased metabolic activity of cells (initially kept for 30 minutes at 4°C) was observed on the 5th and 7th day in cNPSE presence.


Thus, it was found that adding cNPSE and cPSE to the culture medium
of rats’ skin fibroblasts increase the metabolic activity of cells. A dose-dependent effect is observed. The addition of cNPSE to the medium with 2% FBS maintains the metabolic activity of fibroblasts at the level observed in the control samples incubated with 10% FBS. Adding extract also increases the metabolic activity of fibroblasts after hypothermic impact on the cells.

Keywords

fibroblasts; culture; extract; skin; spleen

Hrčak ID:

125480

URI

https://hrcak.srce.hr/125480

Publication date:

31.3.2014.

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