CYTOTOXIC ACTIVITY OF LEUKOCYTES AND BLOOD SERUM OF RATS RELATIVELY TO ALLOGENIC CULTURES OF BONE MARROW CELLS AND PANCREASCELLS

Abstract

Diabetes mellitus is an endocrine disease characterized by a syndrome of chronic hyperglycemia, and is a result of insufficient production and/or action of insulin, which leads to violations of all types of metabolism, primarily carbohydrates, vascular lesions (angiopathy), nervous system (neuropathy) and other organs and systems. Curiosity to study this disease does not go down and encourages researchers to seek new ways of diagnosing and treating diabetes and its complications. Notwithstanding the abovementioned, the main method of treatment of first type diabetes is still using insulin. It should be noted that insulin therapy threatens the development of insulin resistance, the allergic reactions and lipodystrophy. In addition, exogenous insulin injection is not able to prevent the progression of complications of diabetes.

Cell therapy in the diabetes treatment is an alternative method based on using the regenerative potential of stem cells of the adult organism. This is not a new method, more than 30 years ago experimental research on β-cells of the islets of Langerhans transplanting started. However, there still remains a number of unsolved issues on the introduction of cellular technology into practice. One of them is the organism’s interactionwith the transplanted cellular material. We therefore examined the cytotoxic activity of white blood cells and blood serum of rats relatively tothe allogeneic cell material and compared the effects on cells from different organs, bone marrow and pancreas.

Objective: To investigate the cytotoxic activity of white blood cells and blood serum of ratsrelatively to allogeneic cultures of bone marrow cells and pancreascells.

Materials and methods of the research:15non-linear little rats of 12 days of age and 9 non-linear medium male rats weighing 300gr were used in the experiment. Animals were divided into three groups: 1^stis the control one, 2^nd was injected with the bone marrow cells culture, 3^rd was injected with cells culture of the pancreas. All manipulations with animals were performed under general anesthesia for what once was injected intraperitoneally 2% solution of sodium thiopental at the rate of 4mg per 100g of live weight.

On the 15th day blood was taken from sensibilized animals and serum andleukocyteswere isolated from it.

Cytotoxic activity of rat blood serum wasdetermined by serum cultivating together the sensibilized animals’ blood serum, complement and cell culture in standard culture medium.

Cytotoxic activity of white blood cells of rats was determined by cultivating together white blood cells ofsensibilized animals and cell cultures in standard culture medium.

The presence of the cytotoxic effect relatively to the transplanted cells demonstrates the absence of their proliferative activity and reduced viability.

The results of the research: Analysis of the research results showed that the viability and proliferative activity of allogeneic cultures of bone marrow cells and the pancreas in the cytotoxic test with serum and blood lymphocytes of the control and experimental groups of animals probably differ.

We have not remarked the cytotoxicity of serum and blood lymphocytes of animals in the control group as to cultures of bone marrow cells and the pancreas, which is probably due to the lack of prior sensibilization of animal lymphocyte by antigenic determinants of studied animal cells and, therefore, lack of specific antibodies to their receptors in blood serum of animal control group.

15 days after peritoneal injection of cell culture for research animal groups, we have noted that proliferation index of bone marrow cell culture at the 1st passage was 0.9, while in the 4th was greater than in the 1st. The resulting changes coincide with the controlling indicators. These changes showed no cytotoxic influence of serum on culture of bone marrow cells.

We were remarking cytotoxic effect of serum on cell culture of pancreatic, as in the first and fourth passages. This influence ofsensibilizedanimals serum can be explained by heterogeneity of culture and the presence in it of differentiated cells.

Cytotoxic effect of sensibilized animals lymphocytes was not observed on the culture of bone marrow cells and the culture of pancreatic cells as in the first and fourth passages.

Conclusions and prospects:

1. White blood cells of sensibilized animals do not show cytoxic effect on culture cells of the pancreas and bone marrow.

2. Blood serum of sensibilized animals is showing no cytotoxic effect on the culture of bone marrow cells both in the first and fourth passages.

3. A cytotoxic effect of sensibilized animal serum on pancreas cell culture is revealed.

4. Serum cytotoxicity of sensibilized animals decreases with increasing the passage of the pancreas cell culture.