The expression product of VASA gene is widely conserved germ line marker and participates to regulate the development of reproductive system and spermatogenesis in healthy man. Azoospermic is a condition which the man is unable to produce any sperm cells for reproduction activity. This condition produced has bad impact to the man since the ability to produce their own progeny will be interrupted or blocked forever. Not only the family bloodline of the man would be ended, the psychological issue like shamefulness and low self-esteem occurs. In Islam, seeking knowledge is an obligatory to Muslims in order to solve problems and improve lives. Thus to unveil the problem of azoospermic man we aimed to determine the level of VASA gene expression in samples of testes cells of non-obstructive azoospermic (NOA) and compare it with sperms of healthy man. Samples were taken from three NOA patients by testicular sperm extraction (TESE) to obtain testicular biopsies. Testicular cells were isolated and cultured in supplemented knockout DMEM media. VASAgene expression was determined by reverse transcriptase polymerase chain reaction (RT-PCR). The VASA gene expression from sperm of healthy man was also determined for comparison purpose. It was demonstrated that the addition of growth factor significantly increased SSC-like cells colony formation in tissues obtained from NOA patients. No VASA expression was detected in spermatogonial-like stem cells culture on day 1, 7, 14 and 21 in each of the azoospermic samples. Our findings shown VASA gene was not expressed in spermatogenesis in vitroculture that might be associated with the abnormal differentiation of primordial germ cells that lead to male infertility. Islam also teaches us to not have feelings of despair and for problems that we are facing but to find the solution and consider the problem as a test from the Almighty. All the tests should be handled with strong believe since only Allah (SWT) knows what the best for us and each of the tests will have justified wisdom and benefits which we as human being are unable to realise thoroughly.
Azoospermia is present in 15% of infertile cases and it is a major concern due to inability to produce sperm. Most of IVF (in-vitro fertilization) clinics abroad has been using sperm donation via sperm bank facilities as a solution for infertile couple to have their own offspring. In Islam, it is forbidden to use sample from male other than their spouse. It is according to maqasid syari’ah to ensure the heredity of the human being. Based on the latest technology, one approach of stem cell differentiation process had been established to produce mature cells from primitive or immature cells (stem cells). This technology is in line with the concept of maqasid syari’ah since we are using the cells from one person. We try to adopt this technology to study the potential of testicular cells from non-obstructive azoospermic (NOA) patient to undergo in vitrospermatogenesis.Samples were cultured in modified human embryonic stem cells (HESC) media with specific growth factors; basic fibroblast growth factor (bFGF) and leukemia inhibitory factor (LIF). Protein expressions were analyzed by immunofluorescent staining on day 49 and 90 of culture. Results show spermatogonial stem cell-like cells (SSC-like cells) colonies formed after 14 to 21 days, the cells were expanding successfully and were stable for 49 days duration. Then SSCs differentiated into later stage of spermatogenesis on day 90. Four specifics SSCs protein markers were identified on day 49; ITGA1, ITGB1, CD9 and GFRA1 whereas SCP3 and TP1 proteins were expressed on day 90. This in vitrospermatogenesis suggests a possible approach for future for Muslim NOA patients in order to have their own children.
Microalgal lipids are promising and sustainable sources for the production of third-generation biofuels, foods, and medicines. A high lipid yield during the extraction process in microalgae could be influenced by the suitable pretreatment and lipid extraction methods. The extraction method itself could be attributed to the economic and environmental impacts on the industry. This review summarizes the pretreatment methods including mechanical and non-mechanical techniques for cell lysis strategy before lipid extraction in microalgae biomass. The multiple strategies to achieve high lipid yields via cell disruption techniques are discussed. These strategies include mechanical (shear forces, pulse electric forces, waves, and temperature shock) and non-mechanical (chemicals, osmotic pressure, and biological) methods. At present, two techniques of the pretreatment method can be combined to increase lipid extraction from microalgae. Therefore, the extraction strategy for a large-scale application could be further strengthened to optimize lipid recovery by microalgae.