Anti-CD9 monoclonal antibody, clone fCjpTO5

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 vitro spermatogenesis. 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 vitro spermatogenesis suggests a possible approach for future for Muslim NOA patients in order to have their own children.

Herpes simplex virus 1 (HSV-1)-infected cells release extracellular vesicles (EVs) that deliver to uninfected cells viral factors and host components, such as the stimulator of interferon genes (STING), which activates type I interferon upon foreign DNA sensing. The functions of EVs released by HSV-1-infected cells have remained unknown. Here, we describe a procedure to separate the EVs from HSV-1 virions that is based on an iodixanol/sucrose gradient. STING, along with the EV markers CD63 and CD9, was found in light-density fractions, while HSV components accumulated in heavy-density fractions. HSV-1 infection stimulated the release of EVs from the cells. The EVs derived from infected cells, but not from uninfected cells, activated innate immunity in recipient cells and suppressed viral gene expression and virus replication. Moreover, only the EVs derived from infected cells stimulated the expression of a subset of M1-type markers in recipient macrophages. Conversely, EVs derived from STING-knockdown cells failed to stimulate the expression of these M1-type markers, they activated innate immune responses to a lesser extent in recipient cells, and they did not sustain the inhibition of virus replication. These data suggest that STING from the EV donor cells contributes to the antiviral responses in cells receiving EVs from HSV-1-infected cells. Perturbations in the biogenesis of EVs by silencing CD63 or blocking the activity of the neutral spingomyelinase-2 (nSMase-2) increased the HSV-1 yields. Overall, our data suggest that the EVs released from HSV-1-infected cells negatively impact the infection and could control the dissemination of the virus. IMPORTANCE Extracellular vesicles (EVs) are released by all types of cells as they constitute major mechanism of intercellular communication and have the capacity to alter the functions of recipient cells despite their limited capacity for cargo. How the EVs released by HSV-infected cells could alter the surrounding microenvironment and influence the infection currently remains unknown. The cargo of EVs reflects the physiological state of the cells in which they were produced, so the content of EVs originating from infected cells is expected to be substantially different from that of healthy cells. Our studies indicate that the EVs released by HSV-1-infected cells carry innate immune components such as STING and other host and viral factors; they can activate innate immune responses in recipient cells and inhibit HSV-1 replication. The implication of these data is that the EVs released by HSV-1-infected cells could control HSV-1 dissemination promoting its persistence in the host.