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SPERMATOGONIAL STEM CELLS THESIS

Human spermatogonial stem cell clusters were observed after 7 to 10 days in specific culture, then after several passages and successful expanding duration of 52 days, the cells were evaluated by three layer immunocytochemistry test LSAB to stain GPR protein as a surface marker in human spermatogonial stem cells. Discussion Reproductive potential of an organism can be prolonged indefinitely by using germ-line stem cells These integrins act as a laminin receptor and mediate cell attachment to laminin The SSC niche is found dispersed on the basal lamina, which is a two dimensional extracellular matrix along the circumference of seminiferous tubules. 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.

This article has been cited by other articles in PMC. So far, we have been able to culture and proliferate human spermatogonial stem cells in human testicular biopsies successfully by composing some methods such as differential plating, using gelatin and laminin coated plates and SFM that consist human specific growth factors. In addition, in recent decades, in culturing systems laminin coated dishes replaced co-culturing system that based on feeder layers. Reproductive potential of an organism can be prolonged indefinitely by using germ-line stem cells In current study human spermatogonial stem cell were isolated and expanded with the least manipulations in comparison with the other usual isolation methods like florescent or magnetic activated cell sorting.

spermatogonial stem cells thesis

In this way, we get closer to the clinical purpose of spermatogonial stem cell in fertility treatment in the future. 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.

Biopsy samples consisted of at leastand at most 2, cells. Since spermatogenesis does not begin in childhood, freezing mature sperm is impossible.

Preserving male fertility with spermatogonial stem cells

So far, we have been able to thhesis and proliferate human spermatogonial stem cells in human testicular biopsies successfully by composing some methods such as differential plating, using gelatin and laminin coated plates and SFM that consist human specific growth factors.

Before any clinical usage of spermatogonial stem cells, more investigation is required to achieve isolated cancer cells from spermatogonial stem cells.

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spermatogonial stem cells thesis

Then, major of somatic cells will be suspended and transferred to laminin coated dish that xells prepared for specific culture. It should be mentioned that at this stage of studies identifying specific markers, enrichment and isolation undifferentiated spermatogonia from somatic cells and differentiated germ cells were fulfilled by researches.

Isolation and Culture of Human Spermatogonial Stem Cells Derived from Testis Biopsy

Conclusion In current study human spermatogonial stem cell were isolated and expanded with the least manipulations in comparison with the other usual isolation methods like florescent or magnetic activated cell sorting. The migration, self-renewal and differentiation of SSCs was modelled according to xpermatogonial logical parameters provided by researches over the past century.

By considering spermatogonial stem cell number Hela cells as positive control and testis somatic cells derived from an year-old man as negative control are shown in Figure 5. In the current study, after investigation on 12 testicular biopsies, we observed that decrease in the number of somatic cells like myoid and sertoli cells by differential plating thesiis unsuccessful to maintain spermatogonia and early cell death was observed in culture conditions when the number of cells in testicular biopsies was less than 10 6.

In the case of overgrowth of flat cells eclls long extensions which were interpreted as somatic cells, only germ cells and round dividing cells were differentially passed to fresh dishes coated with human placenta laminin by vigorous pipetting Shinohara T, Brinster RL.

This is important because a majority of our prepubertal patients will have a testicular biopsy taken prior to initiation of chemotherapy so we want to make sure there would be no malignant contamination in the sample.

Isolation and Culture of Human Spermatogonial Stem Cells Derived from Testis Biopsy

This work was financially supported by the Avicenna research institute. Human spermatogonial stem cells clusters propagated by expanding the surface area culture after each passage, but the number of fibroblastic cells did not increase as much. This process is based on self-renewal and differentiation of a rare population of the testicular cells called Spermatogonial Stem Cells SSCs.

Towards a knowledge-based human protein atlas. Reproductive potential of an organism can be prolonged indefinitely by using germ-line stem cells The cells were cultured and passaged 6 times during 52 days, in this study.

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Beta 1- and alpha 6-integrin are surface markers on mouse spermatogonial stem cells. Differential plating performed to enrich human spermatogonial stem cells.

Preserving male fertility with spermatogonial stem cells – [email protected]

When the cell number in primary count of testis biopsy was less than 10 6 cells, enriching the spermatogonia was unsuccessful by differential plating. Identification and characterization of repopulating spermatogonial stem cells from the adult human testis.

Introduction Today, treating cancer in children has been improved immensely. The appropriate time to passage the undifferentiated spermatogonia was around the time when morphological changes were taking place.

Progress represented by this thesis research will facilitate translating SSC technologies toward the clinic for preservation and restoration of male fertility.

Preserving male fertility with spermatogonial stem cells Valli, Hanna Preserving male fertility with spermatogonial stem cells. Preserving male fertility with spermatogonial stem cells. We have also studied differential plating to enrich human spermatogonia on digested cell suspension in two aspects.

spermatogonial stem cells thesis

We would like to thank all of the individuals who kindly accepted to participate stej this study. PDF Full Text Restricted up-to 7 May Restricted to Repository staff only Kb Abstract The homeostasis of male genitalia in mammals is maintained by a group of stem cells called spermatogonial stem cells SSCs which replenishes worn out cells in the testicular tissue.

In addition, improved freezing methods is required to maintain the cells for a long period of time. Results indicate that GPRpositive spermatogonia are phenotypically putative human spermatogonial stem cells and retain an undifferentiated status in vitro. So far, human spermatogonial germ cells cluster attached to gelatine or laminin coated plates with the minimum somatic cells accompaniment after several passages Figure 4.