الفهرس 
INTRODUCTIONOnly 14 pages are availabe for public view
 |  | from | 189 |  |  |
| | | from | 189 | | |
Abstract
NOA, particularly of testicular causes, has been recognized as the most severe presentation of male infertility in humans, and its management was, and always will be a challenge for all andrologists around the world.
Most of these cases arise as a side effect of chemotherapy, as chemotherapeutic drugs have shown to be gonadotoxic and able to induce azoospermia in over 90% of cancer male patients.
Busulfan is a chemotherapeutic agent, that is routinely used to induce aplastic anemia in leukemic patients who are waiting for bone marrow transplantation, despite of the fact that, it eradicates the testicular germ cells and causes a long term morphological alteration to the survived spermatozoa.
Stem cells are considered a promising approach for treatment of chemotherapy- induced azoospermia, and stem cells from the amniotic fluid are an attractive and appropriate source for stem cells-based therapeutic preclinical and clinical studies.
The present work aimed at studying the histological changes of the seminiferous tubules of adult male albino rats after induction of azoospermia by busulfan. Furthermore, ensuring homing of the AFSCs in the seminiferous tubules after their transplantation and exploring their possible therapeutic effects.
This study was conducted on 72 albino rats, 10 females aged 6-8 weeks and weighed 150-200 g, and 62 males aged 8-12 weeks and weighed 200-250 g. Female rats were induced to be pregnant by 2 males, to be used as a source for the amniotic fluid, while the remaining 60 male rats were randomly divided as follows;
group I (Control group): 18 rats were subdivided randomly and equally into 3 subgroups: Subgroup IA: 6 rats, each received a single intraperitoneal injection (i.p.) of phosphate-buffered saline (PBS). Subgroup IB: 6 rats, each received a single i.p. injection of dimethyl sulfoxide (DMSO) diluted in PBS (vehicle of busulfan). Subgroup IC: 6 rats, each received a single injection of 1 ml of PBS into the efferent ducts of their testes. Rats of the three control subgroups were euthanized at the end of 28 days.
group II: 42 rats were induced to develop azoospermia. Each rat was treated with a single i.p. injection of busulfan at a dose of 40 ml/kg body weight. After 28 days, the rats were subdivided into 4 subgroups: Subgroup IIA (azoospermia subgroup): 10 rats were euthanized to assess the induced azoospermia. Subgroup IIB (spontaneous recovery subgroup): 10 rats were left with no further intervention for another 28 days, and then they were euthanized, to assess the spontaneous recovery potentials. Subgroup IIC (cell-free medium subgroup): 10 rats received a single injection of 1 ml of cell free-culture medium into the efferent ducts of each testis, and euthanized after another 28 days, to assess the potential of the cell free-culture medium to improve the induced azoospermia. Subgroup IID (AFSCs-treated subgroup): 12 rats were treated with a single injection of the fluorescently-labeled AFSCs suspended in 1 ml of the culture medium, into the efferent ducts of each testis. Two rats of this subgroup were euthanized after 48 hours, for ensuring of homing of the cells in the seminiferous tubules. The remaining 10 rats were euthanized after another 28 days, to assess AFSCs potential to improve the induced azoospermia.
AFSCs were isolated, cultured, passaged, and characterized by:
1. Daily examination of their morphology using the phase contrast inverted microscopy.
2. Colony forming unit (CFU) assay.
3. Flow cytometric analysis of their surface markers.
For their tracking after transplantation, AFSCs were labeled by the fluorescent Cell Tracker CM-Dil, then they were injected into the efferent ducts of rats of subgroup IID. The injected cells survived and homed within the seminiferous epithelium, as detected by the confocal laser scanning (CLS) microscope.