Цитогенетичний аналіз культури клітин жирової тканини щурів на ранніх пасажах

Анотація

CYTOGENETIC ANALYSIS

OF ADIPOSE TISSUE CULTURE IN RATS AT EARLY PASSAGES

 А. I. Mazurkevich, V. V. Kovpak, O. S. Kovpak

 Abstract. Opportunities offered by cell-based technologies in regenerative medicine become more and more appealing not only to experts in molecular biology and genetic engineering but also various medical practitioners. Procedure for obtaining cell cultures is costly, and yet the primary obstacle to cell therapy implementation on a wide scale is its safety and ethical issues. Adipose tissue is an alternative source of adult stem cells. However, a required quantity of cell material may be obtained only through long-term in vitro cultivation. That is why before transplantation it is necessary to study stability of cell culture and to assess risks of neoplastic transformation of cells.

Adipose tissue culture was obtained from subcutaneous fat of outbred rats 4 months old. Cells obtained from passage one to passage six culture were used for cytogenetic analysis. The study was performed taking into account number of cells with abnormal karyotype, cells with micronuclei, binuclear cells and apoptotic cells, and included mitotic count.

In the course of the study we have discovered abnormalities in the genetic apparatus of the cells manifesting themselves in the form of aneuploidy, polyploidy and micronuclei, the number of which varied depending on the passage. However, throughout the cultivation period mutability of the karyotype obtained from adipose tissue of rats did not exceed spontaneous level of mutations typical for this animal species.

According to all studied parameters data concerning genetic stability of adipose tissue culture are within normal range typical for mammals permitting further study of transplantation with minimum risk of neoplastic transformation of the above culture.

Keywords: cytogenetic analysis, micronucleus test, adipose tissue culture