ABSTRACT
In cancer patients, chemo and radiotherapy can cause infertility by damaging spermatogenesis process. This process is based on self-renewal and differentiation of a rare population of the testicular cells called Spermatogonial Stem Cells [SSCs]. Scientists have tried to isolate, enrich and culture Human spermatogonial stem cells, hoping to resolve infertility problems in cancer recovered patients in the future. Spermatogonial stem cells were isolated and purified from human testicular biopsies sample consisting of at least 500,000 and at most 2,000,000 cells. Two enzymatic digestion steps were performed. Enriching methods, differential plating, and specific culture in serum-free medium with added growth factors: human GDNF, bFGF, EGF and LIF was performed on coated dishes. 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 GPR125 protein as a surface marker in human spermatogonial 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. 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
Subject(s)
Humans , Male , Testis , Biopsy , Receptors, G-Protein-Coupled , Cell Culture Techniques , Infertility, MaleABSTRACT
Some cases of male infertility are due to the destructive side-effects of anticancer treatment methods such as chemo and radiotherapies on germ cell lines. The increase in the survival rate of cancer patients who undergo treatment, especially children, has drawn attention to fertility preservation. The most common and effective technique in preserving male fertility is sperm freezing and its subsequent IVF. Children cannot efficiently produce sperm because of their spermatogonial immaturity. One of the strategies to maintain fertility in these patients is to preserve the testes or the germ cells by freezing them for their later maturation and production of fertile sperm, although the state in which the spermatogonia may not undergo maturation is one of the main obstacles faced in this method. Therefore, scientists have attempted to transplant cryopreserved testis tissues or produce in vitro-matured spermatozoa in this group of patients upon anticancer treatment. In this study we reviewed the germ cell biology, the side-effects of chemo and radiotherapies on germ cells and fertility preservation techniques in adults and children undergoing anticancer treatment
ABSTRACT
Current protocols for cancer treatment could lead to the failure of ovarian function and subsequent infertility in women. Therefore, utilizing ways to preserve fertility in these individuals seem to be essential. In this review, the full-text of articles which were accessible and had been published during 1976 to 2009 about different methods of female fertility preservation were collected and studied through various online databases such as PubMed, Science Direct, etc. According to the reviewed articles, there are several methods for fertility preservation in women, including ovarian transposition and oocyte, embryo and ovarian cortex cryopreservation. Ovarian transposition is not useful for preserving fertility in women who undergo chemotherapy. Embryo and oocyte cryopreservations require a delay before starting treatment. Metaphase II oocytes are high-volume and fully-differentiated cells which may sustain injury due to the freezing process restricting the number of collected oocytes and reducing the chances of fertility. On the other hand, ovarian stimulation and oocyte collection are not practical in young patients, especially in underage girls. In addition to the restrictions on the number of collected embryos and the raised legal and ethical issues, embryo crypreservation is limited to adults and married women. In comparison to other methods, cryopreservation of the ovarian cortex seems to be more appropriate as ovarian tissue is resistant to cryopreservation and it is easy to be collected by laprascopy, making it practical for use in premature girls. Furthermore, the large number of follicles in the ovarian tissue increases the chances of fertility preservation in women. In general, several parameters including the type, time and duration of treatment, cancer type, age and marital status determine the efficacy of each method
ABSTRACT
Spermatogonia are the male germ line stem cells whose life long expansion is needed for permanent production of spermatozoa. The present study was designed to examine the effect of hCG treatment on germ cell proliferation following stem cell transplantation in mice. Spermatogonial stem cells were isolated from neonatal mice testes and characterized by alkaline phosphatase, immunoreactivity and morphological analysis. hCG was injected into normal and cell transplanted mice. We then evaluated the testosterone levels and cell number in normal mice. After that, cyclin B1 gene expression was investigated in transplanted mice. Different doses of busulfan were injected to investigate the effects of chemotherapy on morphological criteria and preparation of recipient mice for transplantation. In this report we show proliferative potential of spermatogonial stem cells after cytotoxic treatment, transplantation efficiency by semi-quantitative RT-PCR, and hCG effect on stem cell regeneration in normal mice and following cell transplantation. The results indicate the spermatogonial stem cells can proliferate after transplantation, and the efficiency of their transplantation depends on hormonal treatment. Therefore, hormonal treatment after stem cell transplantation will be a powerful avenue for increasing the efficiency of transplantation and fertility restoration