However, these options are not available to all patients, especially the prepubertal patients who are not yet producing mature gametes. Author information Article notes Copyright and License information Disclaimer. On the other hand, the number of spermatogonial stem cells in one testis biopsy is not sufficient to transplant for healing the damages of germinal layer caused by cancer therapy. 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. 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. He et al , isolated GPRpositive spermatogonia from adult human testis, followed by magnetic-activated cell sorting.

In Chapter 3, I used this knowledge about the phenotype of human spermatogonia to show that the best method to cryopreserve intact human testicular pieces is controlled slow-freezing. Mohammad Mehdi Akhondi, Ph. 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 was 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 current study, we tried to isolate, enrich and specifically culture spermatogonial stem cells derived from tiny biopsy samples. The colonies condition was not mentioned for different cases on gelatin and laminin coated plated when the primary count was less than 10 6 cells in biopsy samples.

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

Author information Article notes Copyright and License information Disclaimer. In recent years, three major requirements needed to advance spermatogonial stem cell research were i an in vivo assay for stem cell function like methods for spermatogonial transplantation, ii knowledge of stem cell markers, and iii a method to maintain the stem cells continuously in culture Similarly, division and differentiation was also controlled by density stress through various thresholds.


Preserving male fertility with spermatogonial stem cells

Pluripotent stem cells derived from adult human testes. In mouse, these cells are housed by specialised spermatogonlal structures called seminiferous tubules. A MatlabTM program was developed to simulate the behavior of SSCs in their niche within the seminiferous tubules with logical rules. In contrast to the other SSCs isolation and culture methods, this system is based on the testicular biopsies against large samples, thus suggested method in this study is closer to clinical usage in the future.

With these metrics in concern, a biopolymer scaffold was prepared by using spermatogonlal to mimic the testicular tissue.

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

Then, dehydration was performed through a series of graded alcohols. Gelatin is a derivative of matrix constituency and assists in attachment of cells to a surface and allows them to proliferate Human spermatogonial stem cells clusters propagated by expanding the surface area culture after each passage, but the number of fibroblastic cells did not increase spermayogonial much.

For Immunochemistry in those cases, it has been suggested to replace three layers staining instead of two layers staining Just by morphology, human spermatogonial stem cell clusters condition was not so different on gelatin or laminin during several passages. In cancer spfrmatogonial, chemo and radiotherapy can cause infertility by damaging spermatogenesis process.

Received Apr 18; Accepted Sep Spetmatogonial are also deeply indebted to the personnel of the Avicenna Reproductive Biotechnology and Nanobiotechnology departments. Kanatsu-Shinohara et al cultured SSCs in such a way that these cells propagated themselves, while retaining their capacity to repopulate a recipient mouse testis tsem transplantation 8.

Discussion Reproductive potential of an organism can be prolonged indefinitely by using germ-line stem cells Nevertheless, isolating germ cells from blood-related malignancies needs more investigation before any clinical procedures can take place.


Reconstitution of spermatogenesis from frozen spermatogonial stem cells. Oncofertility fertility preservation for cancer survivors. Beta spermatogoniall and alpha 6-integrin are surface markers on mouse spermatogonial stem cells. The cells were cultured and passaged 6 times during 52 days, in this study. Avicenna J Med Biotechnol.

spermatogonial stem cells thesis

Spermatogonial stem cells clusters were passaged every days. The specific culture of human spermatogonial stem cells was evaluated and approved by collecting the cultured cells after 52 days and DAB staining GPR as specific surface marker on these cells.

In this way, we could prevent excessive loss of cells and restrict proliferation somatic cells by decreasing serum in culture condition at last. The accurate comparison between spermatogonial stem cell clusters condition on laminin or gelatin coated plates would be reliable by following cellss fate of the cells after transplantation.

spermatogonial stem cells thesis

Isolation of male germ-line stem cells; influence of GDNF. At the slermatogonial, a somatic cell and at the right, a human germ cell colony are shown. Slides were fixed in NBF for 10 min.

Then it is likely, that the cells could not interact with each other and early cell death would be observed. 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. The progenitor Adark-spermatogonia apparently are stems or reserve spermatogonia 2. Since spermatogenesis does not begin in childhood, freezing mature sperm is impossible. Seandel et alhave identified GPR as a surface marker for self renewing clonogenic spermatogonial progenitor cells in mouse 5.