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Male causes of infertility constitute approximately half of
infertility cases. Furthermore, cases of non-obstructive
azoospermia (NOA) represent 10% of male infertility and have no
definite effective treatment. Recently, stem cells have been
suggested to treat different conditions. Adipose derived
mesenchymal stem cells (ADMSCs) proved to be more
convenient than bone marrow derived mesenchymal stem cells
(BMMSCs). They could be extracted from different sources with
minor invasive techniques, proliferate in less time and give more
Animal models of NOA were induced by intratesticular
CaCl2 injection. It causes irreversible chemical castration with
minimal or no side effects. Accordingly, the current study was
designed to evaluate the therapeutic role of (ADMSCs) in
treatment of NOA induced by CaCl2.
Forty five adult male albino rats of average weight of 225
gm were used in the current study. All rats were given oral
paracetamol (1ml syrup) (before the injection). They were divided
into four groups:
group I: It included fifteen rats. Ten rats (subgroups Ia, Ib) were
injected with a single intra-testicular dose of 0.1 ml saline/100gm
Subgroup Ia: five rats were sacrificed after one week.
Subgroup Ib: five rats were sacrificed after nine weeks. Subgroup Ic: five rats were sacrificed for the obtaining of
adipose tissue derived mesenchymal stem cells (ADMSCs).
group II: It included ten rats. Each rat was subjected to a single
intra-testicular injection of calcium chloride (CaCl2)
(2.5mg/testis/100gm body weight) dissolved in 0.1 ml saline/
testis. The animals were sacrificed after one week.
group III: It included ten rats. They were given CaCl2 as in
group II. After one week from CaCl2 injection, 100 𝜇L of
ADMSCs mixture (106 cells) dissolved in 0.1 ml saline/ testis was
injected intra-testicular. The animals were sacrificed eight weeks
after stem cells injection.
group IV: It included ten rats. Each rat was given CaCl2 in the
same manner as in group II and sacrificed after nine weeks to be
compared with group III.
At the designated times, the animals were sacrificed by
cervical dislocation. The testes were dissected out and processed
for both light (H&E, PAS, Masson trichrome and Toludine blue
stains) and electron microscopic examination. Sperm count
smears sections preparation and total testosterone hormonal assay
were done. Morphometric and statistical analysis were performed.
The results of this study revealed spermatogenic epithelium
degeneration with various degrees of apoptosis and necrosis in the
seminiferous tubules of group II. Sloughing of epithelial lining of
the seminiferous tubules with formation of necrotic tissue was
detected. Damaged Leydig cells, inflammatory cells and
multinucleated giant cells were also observed in group II.
Injection of ADMSCs in group III resulted in great
improvement of the structure of the testis being comparable to the control group. The seminiferous tubules showed well organized
spermatogenic epithelium and sperms in the lumen.
However, group IV showed a similar picture to that of
group II with more damage of the testicular tissue and extensive
collagen fibers deposition. Apoptosis and necrosis were obvious
in this group with presence of damaged Leydig cells, chronic
inflammatory cells (macrophages) in the interstitium.
Statistically, groups II and IV showed a significant
decrease in the mean sperm count and the serum testosterone
level as compared to the control group. On the other hand, group
III showed a non-significant change as compared to the control
from the previous data, it was concluded that ADMSCs
injection resulted in great improvement and regeneration of the
structure of the testis following the testicular damage induced by
NOA CaCl2 model.