SPERMATOGONIAL STEM CELLS THESIS

Gelatin has often been used to coat plastic surfaces and enhance attachment of various cell type experiments in vitro culture. The tubular nature of the scaffold mimicked seminiferous tubules in dimension. Then, dehydration was performed through a series of graded alcohols. The behavior of SSCs in their niche was assumed to be dependent on cell density in that niche area. Isolation and culture of human spermatogonial stem cells was fulfilled from TESE positive testicular biopsies with at least , to at most 2,, cells. The model predicted that there was 12 Asingle cells, 15 Apaired cells and 19 Aaligned cells per Sertoli cells in the niche of mouse. In the case of overgrowth of flat cells with 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

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. Primary spermatogonial germ cells cluster was near or on the somatic cells Figure 2. Towards a knowledge-based human protein atlas. Isolation of male germ-line stem cells; influence of GDNF. In the cases that biopsies contained less than 10 6 cells, we bypassed the differential plating method that was mentioned above. Different types of human spermatogonia include progenitor Adark-spermatogonia, progenitor Apale-spermatogonia, committed Apale-spermatogonia, and B-spermatogonia. The behavior of SSCs in their niche was assumed to be dependent on cell density in that niche area.

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On the other hand, we observed that when primary cell count was more than 10 6 cells in digested cell suspension, differential plating, was beneficial to enrich human spermatogonia and decrease disturbing of somatic cells before transferring the cells to SFM. We are wpermatogonial deeply indebted to the personnel of the Avicenna Reproductive Biotechnology and Nanobiotechnology departments.

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

Differential plating performed to enrich human spermatogonial stem cells. Today, treating cancer in children has been improved immensely.

The SSC niche is found dispersed on the basal lamina, which is a two dimensional extracellular matrix along the circumference of seminiferous tubules. Isolation and culture of human spermatogonial stem cells was fulfilled from TESE positive testicular biopsies with at leastto at most 2, cells.

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Materials and Methods Sampling and digestion 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 However, majority of thesi knowledge about spermatogonial stem cells SSCs comes from rodents and not much is known about humans. In mouse, these cells are housed by specialised tubular structures called seminiferous tubules.

The model predicted that there was 12 Asingle cells, 15 Apaired cells and 19 Aaligned cells per Sertoli cells in the niche of mouse. In general, when spermatogonial stem cells have been separated by conjugated antibodies, a high degree of cell purity was obtained, but a low number of cells resulted. At the left, a somatic cell and at the right, a human germ cell colony are shown.

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Enriching methods, differential plating, and specific culture in serum-free medium with added growth factors: Clels Center Support Center. Primary spermatogonial germ cells cluster was near or on the somatic cells Figure 2. Reconstitution of spermatogenesis from frozen spermatogonial stem cells. The model outcome was validated with literature.

Preserving male fertility with spermatogonial stem cells

The second question asks the appropriate time to gain successful attachment of human sertoli and myoid cells to plastic dishes. The ULS Office of Scholarly Communication and Publishing fosters and supports new modes of publishing and information-sharing among researchers.

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.

The migration, self-renewal and differentiation of SSCs was modelled according to the logical parameters provided by researches over the past century.

spermatogonial stem cells thesis

In the case of overgrowth of flat cells with 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 After the first passage, the clusters got more typical appearance. Human germ cells colonies were attached to laminin coated plate without fibroblast accompaniment after the second passage.

After 3 hours examination, we observed many cells that had suspended and the majority of cells transferred with spermatogonia to SFM.

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

Nevertheless, the plate that related to overnight primary culture showed appropriate somatic cells attachment during this period Figure 1B. The progenitor Adark-spermatogonia apparently are stems or reserve spermatogonia 2. A MatlabTM program was developed to simulate the behavior of SSCs in their niche within the seminiferous tubules with logical rules. 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.

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spermatogonial stem cells thesis

Spermatogenesis following male germ-cell transplantation. Thesie Isolation and culture of human spermatogonial stem cells was fulfilled from TESE positive testicular biopsies with at leastto at most 2, cells.

Differential plating was suggested to enrich spermaogonia in cell suspension derived from testis sample after enzymatic digestion by some articles 9 Kinetics of spermatogenesis in mammals: Introduction Today, treating cancer in children has been improved immensely.

spermatogonial stem cells thesis

Abstract Background In cancer patients, chemo and radiotherapy can cause infertility by damaging spermatogenesis process. Just by morphology, human spermatogonial stem cell clusters condition was not so different on gelatin or laminin during several passages.

Assessment of spermatogobial gradient on enrichment and culture of perinatal porcine spermatogonial stem cells; [NCSU libraries]: