Animal Hormonal Status Changes in Androgen Deficiency (AD) Settings under Influence of Stem Cells Syngeneic Culture. Cellular Tracking and Fluorescence Imaging ex vivo/in vivo

The age-related male hypogonadism is one of the most common causes of infertile marriage in Ukraine. The pathogenesis of this condition is bound to decrease of the level of testosterone (T), the main masculine hormone. The peak levels of T observed in males 25 – 30 years of age. According to MMAS data the general T concentration declines by 0.8% per year starting from 30 – 35 years of age, while biologically accessible T level decreases at the rate of 2 – 3 % annually. As such, it would be logical to consider that treatment is the restoration of required hormone level. Traditionally the correction is achieved by hormone replacement therapy (HRT). However, any kind of replacement pharmacotherapy also assumes the number of limitations, and potential for adverse events development. Additionally, there is an inconvenience of permanent medications intake. The T-therapy demands the strict adherence to certain conditions and precautions. The above stated clearly indicates the necessity of search for alternative ways of AD correction. The most popular method is an application of bone marrow stromal cells (BMSC), since they can be harvested from adult individual for further autotransplantation. Our experimental work describes the local transplantation of syngenenic cells in rats. We conducted a number of experiments on rats reproducing at first the AD model by administration of CdCl2, which was permanent and irreversible without therapy. We assessed the hormonal status in animals within the experiment, as well as conducted the tracking of involved BMSC marked with green fluorescent protein (GFP). 28 days upon single injection of toxin in the dose of 150 mg/kg of weight the following changes in animals' hormonal status were observed: the rate of T-level decline decreased; the FSH, LH and GnRH levels increased; the improvement of free androgen index (FAI). The bilateral intratesticular injection of 200 000 cells resulted in the most positive outcome: the T-level increased by 55.1% comparing to experimental model of disease; the FSH level decreased by 37.5%, the LH level decreased by 18.3% and the GnRH level decreased by 37,4%; FAI increased by 97.2%. The comparing of the results obtained with those in the intact group revealed the following: T-level was slightly lower by 3.4% than in the intact group; the FSH level was 2.5% lower than in the intact group; the LH levels were equal for both groups; the GnRH level was 10.8% lower than in the intact group; FAI was 13.6% higher than in the intact group. The study of tracking of GFP-marked BMSC both ex vivo and in vivo revealed that cells spread equally along their intratesticular inoculation. The assessment was done immediately after BMSC injection ex vivo, and on Day 1 and Day 28 of the experiment in order to confirm cells presence in the testicles of animals under experiment. The conducted experiments prove that bilateral intratesticular BMSC injection in the dose of 200 000 per each testicle results in the most positive outcome in correction of experimental Age-related Androgen Deficiency model. We created the cell bank of BMSC with GFP expression which can be used for the research of tracking of cells in testicles. Our cell culture can be utilized in applied methods of fluorescence imaging at the time of transplantation. Also, it was demonstrated that after injection in vivo the cells migrate into intercanallicular space of the testicles and spread in there equally maintaining the vitality for up to 28 days.