Application of cultivation for experimental impact assesment of fetal neurogenic cells’ supernatant on C6 glioma cells


  • L. D. Lyubich
  • V. M. Semenova
  • L. P. Stayno
Keywords: rat progenitor neurogenic cells, supernatant, rat glioma C6, cytotoxic index, mitotic index

Abstract

Introduction. Despite intensive research of malignant gliomas in the world, today significant progress in their treatment are not attained by invasiveness and high retsydyvnist gliomas. The combination in treatment of cancer surgery, radiation and chemotherapy is the gold standard, but does not ensure its effectiveness. One alternative approaches to solve this problem is to use neurogenic stem cells and progenitor cells.

It is known that neural progenitor cells exhibit antitumor properties in mice and rats. Multipotent neural progenitor cells of human, rat and mouse expressing both proinflammatory and suppressor cytokines. However, the mechanism of antitumor properties of neurogenic progenitor cells remains unclear.

According to previous studies, fetal nerve cells of rats 18-20 days gestation exhibit antitumor effect in experimental glioma cells in vitro and 101.8 in vivo at heterotransplantatsiyi joint capsule in the kidneys of mice. Shown anti-tumor effect of the supernatant of fetal rat brain cells in rats with glioma 101.8 in vivo.

The purpose of the study was to evaluate the effect of rat fetal neurogenic cells supernatant (RFNS) on cultured cells of rat glioma C6.

Materials and methods. RFNS from suspensions of neurogenic cells of rat brain of 9-th (E9) and 14-th (T14) days of gestation (0,01, 0,10 mg/ml) were added tо cell suspensions of rat glioma C6; after 24 h of incubation cell suspensions were analyzed for number of viable cells, cytotoxic index (CІ) was calculated. RFNS (0,01, 0,10 mg/ml) were added tо primary cultures of rat glioma C6, after 24 and 48 h of incubation cytological preparations were analized and mitotic index was determined.

The supernatant neural cells (NCC) received suspensions of neurogenic rat brain cells at 9 (E9) and 14 (E14) day of gestation.

Results. Supernatants fetal rat brain both studied gestation (E9, E14) revealed a cytotoxic effect on all samples C6 glioma cells in short-term cultures. The level of exposure cytotoxic intensified with increasing duration of incubation of cells with supernatants. With increasing concentration of NCC (E9) from 0.01 to 0.10 mg/ml in the suspensions THESE C6 glioma cells increased and reached (37,61±2,57)% and (50,12±4,55)%, respectively, at 24 hours. and 48 hours. incubation.

At 4-5 day culture experienced a gradual rozryhlennya mikroeksplantativ cell to form a monolayer of tumor cells rozroschen unipolar, triangular or rhomboid shape with fairly long spikes. In these areas crop growth zone were more common areas sitkopodibnoyi architectonics due to the formation of intercellular communications.

In subsequent observation period (8-9 days) in cultures of C6 glioma There was a further dilution of cell arrays zone growth due to spontaneously share desquamation of dead cells appeared significant number of apoptotic cells, reflecting the spontaneous death of cultured cells. Necrobiotic processes spontaneous death covered the vast number of cells zone growth remained preserved some areas sitkopodibnyh structures and cellular sferoyidni microaggregates, surrounded by a monolayer of epithelial cell growths undifferentiated type.

Conclusions.

1. The supernatant neural cells fetal rat brain different terms of gestation and antymitotychnyy found cytotoxic effects on cultured C6 glioma cells, which intensified with increasing gestational age rat brain, lengthening the duration of incubation was dose-dependent.

2. Primary cultures of rat brain gliomas is adequate experimental model for evaluation of antitumor action neurogenic cells of fetal rat.

3. Using primary cultures of rat brain gliomas to evaluate individual sensitivity of tumor cells to the studied biological product.

Published
2017-10-27
How to Cite
Lyubich, L. D., Semenova, V. M., & Stayno, L. P. (2017). Application of cultivation for experimental impact assesment of fetal neurogenic cells’ supernatant on C6 glioma cells. Reports of Morphology, 21(2), 352-357. Retrieved from https://morphology-journal.com/index.php/journal/article/view/167
Section
ORIGINAL INVESTIGATIONS