The influence of the conditioned environment of sheep mesenchymal stromal cells on the qualitative parameters of sperm cells

Abstract

The purpose of this research was to study the influence of the conditioned medium of mesenchymal stem cells from adipose tissue and bone marrow of sheep on the quality parameters of sperm during a threehour incubation. The sperm of sexually mature rams of the Romanov and Dorper breeds at the age of 1-2.5 years was taken and examined. A total of 10 samples were taken (n=10). Conditioned medium (CM) was collected after 72 h of cultivation in mattresses with 90-100% cell monolayer. Three groups of samples, 10 in each, were formed: control (spermatozoa 7x1010 cells/ml + PBS buffer 100 μl, pH – 7.4), first experimental (spermatozoa 7x1010 cells/ml + conditioned medium (CM) MSC BM 2x108 cells/ml, pH – 7.2), the second experimental one (spermatozoa 7x1010 cells/ml + conditioned medium (CM) MSCs in adipose tissue 2x108 cells/ml, pH – 7.2). Comparisons of values were made at each incubation stage (0, 1, 2 and 3 hours) within each group. It was found that the use of conditioned medium of mesenchymal stem cells from adipose tissue and bone marrow of sheep at a concentration of 2x108 cells/ml and spermatozoa at a concentration of 7x1010 cells/ml in terms of improving the quality indicators of germ cells. During a three-hour incubation of sheep sperm with conditioned medium of mesenchymal stem cells (MS), a decrease in the number of progressively moving sperm was observed. At the same time, a statistically significant difference was recorded in the first experimental group (CS MSC BM) on the 3rd hour of incubation: 1.9 times (p<0.01), as well as in the second experimental group (CS MSC VT) on the 2nd and 3rd hour of incubation compared with indicators of 0 hour of incubation: 1.4 times (p≤0.05) and 2.4 (p<0.01), respectively. A statistically significant decrease in the number of morphologically normal sperm was recorded during an hour-long incubation with conditioned medium of MSCs from sheep bone marrow and amounted to 43.90±2.22% (p=0.04).