1.Spermatogonial Stem Cells Protein Identification in In Vitro Culture from Non- Obstructive Azoospermia Patient
Azantee Yazmie Abdul Wahab ; Muhammad Lokman Md. Isa ; Roszaman Ramli
Malaysian Journal of Medical Sciences 2016;23(3):40-48
Background: Spermatogonial stem cells (SSCs) are classified as a unique adult stem cells that have capability to propagate, differentiate, and transmit genetic information to the next generation. Studies on human SSCs may help resolve male infertility problems, especially in azoospermia patients. Therefore, this study aims to propagate SSCs in-vitro with a presence of growth factor and detect SSC-specific protein cell surface markers. Methods: The sample was derived from non-obstructive azoospermic (NOA) patient. The disassociation of SSCs was done using trypsin. Specific cultures in serum-free media with added basic fibroblast growth factor (bFGF) were developed to support self-renewal division. This undifferentiated protocol was performed for 49 days. Cells were analysed on days 1, 7, 14, 21, and 49. Results: Human SSCs began to aggregate and form colonies after 14 to 21 days in specific culture. Then, the cells were successful expanded and remained stable for a duration of 49 days. Four specifics markers were identified using immunofluorescence in SSCs on day 49: ITGα6, ITGβ1, CD9, and GFRα1. Conclusion: This approach of using in vitro culture with additional growth factor is able to propagate SSCs from non-obstructive azoospermia patient via detection of protein cell surface markers.
2.Ameliorative Effects of Aquilaria malaccensis Leaves Aqueous Extract on Reproductive Toxicity Induced by Cyclophosphamide in Male Rats
Redzuan Nul Hakim Abdul Razak ; Faridah Ismail ; Muhammad Lokman MD Isa ; Azantee Yazmie Abdul Wahab ; Hussin Muhammad ; Roszaman Ramli ; Raja Arif Shah Raja Ismail
Malaysian Journal of Medical Sciences 2019;26(1):44-57
Background: Cyclophosphamide (CP) is a widely used anti-neoplastic and
immunosuppressive agent that is associated with adverse side effects including reproductive
toxicity. Aquilaria malaccensis (AM) is a traditional medicinal plant which was reported to exhibit
high anti-oxidant and free radical scavenging properties. The present study was aimed to evaluate
the protective effects of AM leaves extract on sperm quality following toxic exposure to CP.
Methods: Forty-eight male Sprague Dawley rats were allocated into eight groups of six
rats (n = 6): control, CP only (200 mg kg−1), AM only (100 mg kg−1, 300 mg kg−1 and 500 mg kg−1)
and CP + AM (100 mg kg−1, 300 mg kg−1 and 500 mg kg−1). Animals were sacrificed after 63 days of
treatment and the sperm from the caudal epididymis was taken for sperm analysis.
Results: The body and the reproductive organs weight, sperm count and motility did not
differ between CP and other groups (P > 0.05). A significant increase (P < 0.05) in percentage of the
dead and abnormal sperm were seen in the CP alone treated group compared to the control group.
Co-administration of AM to the CP exposed rats significantly reduced the (P < 0.05) percentage of
abnormal sperm as compared to the CP only group.
Conclusion: Overall, the present results represent the potential of AM to protect against
CP induced reproductive toxicity.