SPERMATOGONIAL STEM CELLS THESIS
For the first time, Dirami et al performed differential plating to enrich porcine Type A Spermatogonia to decrease Sertoli and myoid cells 9. Abstract Background In cancer patients, chemo and radiotherapy can cause infertility by damaging spermatogenesis process. For these patients, there are several stem cell technologies in the research pipeline that may give rise to new fertility options and allow infertile patients to have their own biological children. Methods Spermatogonial stem cells were isolated and purified from human testicular biopsies sample consisting of at least , and at most 2,, cells. Two enzymatic digestion steps were performed by 2.
Nevertheless, the plate that related to overnight primary culture showed appropriate somatic cells attachment during this period Figure 1B. Discussion Reproductive potential of an organism can be prolonged indefinitely by using germ-line stem cells In Chapter 4, I used the phenotype of human spermatogonia from Chapter 2, to show that it is possible to separate potentially therapeutic human spermatogonial stem cells from malignant contamination. Spermatogonial stem cells are the foundation of spermatogenesis and may have application for preserving and restoring male fertility. After the first passage, the clusters got more typical appearance. Abstract Background In cancer patients, chemo and radiotherapy can cause infertility by damaging spermatogenesis process.
Shinohara T, Brinster RL.
Two enzymatic digestion steps were performed by 2.
Mohammad Mehdi Akhondi, Ph. In the next step, after expanding human spermatogonial stem cells, we evaluate our culture system by immunocytochemistry test to stain GPR protein as surface marker in human spermatogonial stem cells. They are the only cell type that can tjesis and restore fertility to congenitally infertile recipient mice following transplantation 13 In the cases that biopsies contained less than 10 6 cells, thseis bypassed the differential plating method that was mentioned above.
The spermatogonial cells, which remained in suspension, were collected before plating 9. A MatlabTM program was developed to simulate the behavior of SSCs in their niche within the seminiferous tubules with logical rules.
This study is based on 12 biopsy samples taken from different azoospermic patients referred to Avicenna infertility clinic center for operating TESE procedure testicular sperm extraction The model predicted that there was 12 Asingle cells, 15 Apaired cells and 19 Aaligned cells per Sertoli cells in the niche of mouse.
For this purpose, frozen year-old man testis, was thawed. After reviewing previous experiments using these technique on testicular tissue, the effect of viability on subsequent experiments is still questionable since viability was not often reported The cells were cultured and passaged 6 times during 52 days, in this sprrmatogonial.
Preserving male fertility with spermatogonial stem cells
For these patients, there are several stem cell technologies in the research pipeline that may give rise to new fertility options and allow infertile patients to have their own biological children.
In culture field, Kanatsu-Shinohara et al cultured mouse SSCs in such a way that these cells propagated themselves on a feeder layer. Support Center Support Center. NC State University; Acknowledgement This work was financially supported by the Avicenna research institute.
Isolation and Culture of Human Spermatogonial Stem Cells Derived from Testis Biopsy
In this way, we could culture human spermatogonial stem cells clusters derived from tiny testis biopsy. However, chemotherapy and radiotherapy as the methods of cancer treatment damage different types of cells especially proliferating cells like spermatogonial ones in testis.
In the current study, we tried to isolate, enrich and specifically culture spermatogonial stem cells derived from tiny biopsy celle. Then, major of somatic cells will be suspended and transferred to laminin coated dish that was prepared for specific culture.
During this time, spsrmatogonial and sertoli cells make opportunities to attach culture dishes. Culture conditions and single growth factors affect fate determination of mouse spermatogonial stem cells. Beta 1- and alpha 6-integrin are surface markers on mouse spermatogonial stem cells.
Hela cell line was used as positive control After overnight incubation, floating cells were collected and cultured in uncoated dishes with supplemented StemPro with human growth factors. Hence, before each treatment, the possibility of specific isolation, enrichment, culturing and expanding tjesis stem cells as major sperm generators should be provided to autotransplant and recover fertility after cancer treatment.
This is important because a majority of our prepubertal stej will have a testicular biopsy taken prior to initiation of chemotherapy sttem we want to make sure there would be no malignant contamination in the sample.
Decreasing the attaching cells during differential plating makes the confluency of cells in vitro condition too little. The plate that related to 3 hr primary culture showed little cell attachment.
Immunochemistry Culture of human spermatogonial stem cells was evaluated by immunocytochemistry test. After performing clinical procedure and confirming active spermatogenesis, and obtaining verbal informed consent from the patients, surplus testis samples were donated to the research laboratory. This work was financially supported by the Avicenna research institute.