Affiliation:
1. State Institution «Republican Scientific and Practical Center for Transfusiology and Medical Biotechnologies», Ministry of Health of the Republic of Belarus, Minsk, Belarus
2. State Institution "Republican Scientific and Practical Center of Transfusiology and Medical Biotechnologies" Ministry of Health the Republic of Belarus
3. State Institution «Republican Scientific and Practical Center for Transfusiology and Medical Biotechnologies», Ministry of Health of the Republic of Belarus, Minsk, Belarus.
4. Belarusian State Medical University.Minsk, Belarus
5. Belarusian State Medical University, Minsk, Belarus
Abstract
One of the important but insufficiently studied feature of mesenchymal stromal cells (MSCs), their microvesicles (MV MSCs) and plasma enriched with soluble platelet factors (PRP) is their trophic function, which ensures the viability of target cells.
The purpose of this study is to analyze the capability of MSC, MV MSC, PRP to provide effect on viability, spontaneous and activation-induced apoptosis of rat spleen lymphocytes during culturing in vitro.
Methods: cell culture, flow cytometry.
Results: The presence of MSCs at the concentrations of 10% and 20% caused an increase in the number of living intact and PMA-activated lymphocytes by the end of 3-day in vitro cultivation. Compared to control, the amount of necrotic cells was 8.3-13.5 fold decreased, the number of apoptotic cells was 2.3-4 fold deceased, mainly due to lymphocytes at late stage apoptosis. MSCs microvesicles at the concentrations used did not reveal significant impact on the viability of lymphocytes cultured in vitro but reduced the level of apoptosis of intact and PMA-activated lymphocytes by 3,6 (р=0,03) and 4,8-5,2 (р=0,048, р=0,03) fold respectively. Studies with rat PRP have shown that at a concentration of 1.25% it has a growth-stimulatory activity against MSCs, but not lymphocytes cultured in vitro. In the culture of intact lymphocytes, PRP had no significant effect on cell viability with a slight decrease (2.7–2.9 fold, p 0.05) the number of necrotic cells. In the culture of PMA-activated lymphocytes, 1.25–2.5% PRP provided an increase in the amount of living cells by 1.6-2.2 fold (р=0,002, р=0,01) and a 2-fold decrease (р=0,02) in the number of necrotic cells.
Conclusion. MSCs, microvesicles and plasma enriched with soluble platelet factors in decreasing order cause rat spleen lymphocytes enhanced viability and suppressed late apoptosis during in vitro cultivation. Lymphocytes activated by forbol myristate acetate are more sensitive to their vital action compared to resting cells.