ABSTRACT
An ideal animal model of azoospermia would be a powerful tool for the evaluation of spermatogonial stem cell (SSC) transplantation. Busulfan has been commonly used to develop such a model, but 30%-87% of mice die when administered an intraperitoneal injection of 40 mg kg-1. In the present study, hematoxylin and eosin staining, Western blot, immunofluorescence, and quantitative real-time polymerase chain reaction were used to test the effects of busulfan exposure in a mouse model that received two intraperitoneal injections of busulfan at a 3-h interval at different doses (20, 30, and 40 mg kg-1) on day 36 or a dose of 40 mg kg-1 at different time points (0, 9, 18, 27, 36, and 63 days). The survival rate of the mice was 100%. When the mice were treated with 40 mg kg-1 busulfan, dramatic SSC depletion occurred 18 days later and all of the germ cells were cleared by day 36. In addition, the gene expressions of glial cell line-derived neurotrophic factor (GDNF), fibroblast growth factor 2 (FGF2), chemokine (C-X-C Motif) ligand 12 (CXCL12), and colony-stimulating factor 1 (CSF1) were moderately increased by day 36. A 63-day, long-term observation showed the rare restoration of endogenous germ cells in the testes, suggesting that the potential period for SSC transplantation was between day 36 and day 63. Our results demonstrate that the administration of two intraperitoneal injections of busulfan (40 mg kg-1 in total) at a 3-h interval to mice provided a nonlethal and efficient method for recipient preparation in SSC transplantation and could improve treatments for infertility and the understanding of chemotherapy-induced gonadotoxicity.
Subject(s)
Animals , Male , Mice , Adult Germline Stem Cells/transplantation , Azoospermia/chemically induced , Busulfan/toxicity , Disease Models, Animal , Infertility, Male/chemically induced , Injections, Intraperitoneal , Spermatogenesis/drug effects , Spermatogonia/drug effects , Stem Cell Transplantation/methodsABSTRACT
Background@#The increasing number of young survivors after cancer treatment and of patients with non-malignant conditions who are at risk for subfertility has resulted in a demand for fertility preservation services, including the Philippines.@*Objective@#The aim of this paper is to provide an overview of the history, indications, and management principles of fertility preservation. Also, the available strategies in the Philippines in both pre-pubertal and post-pubertal men and women and future directions of the field in the country will be discussed.@* Materials and methods@#Literature review, historical accounts@*Results and conclusions@#Fertility preservation should be a priority when treating children and adults of reproductive age with agents that have deleterious effects on the gonads. If harmful treatment will be used, the options of fertility preservation should be discussed, as early as possible by the primary physician in collaboration with the oncologist and the reproductive medicine specialist. Most of the known options for fertility preservation are available in the Philippines and are being implemented in the local IVF centers. Recent developments hint of a potentially faster progress in the field with the establishment of the Philippine Society for Fertility Preservation in collaboration with other professional societies and a linkage with the Department of Health with the signing into law of the National Integrated Cancer Control Act of 2019.
Subject(s)
Fertility Preservation , Cryopreservation , Oocytes , Ovary , FertilityABSTRACT
An ideal animal model of azoospermia would be a powerful tool for the evaluation of spermatogonial stem cell (SSC) transplantation. Busulfan has been commonly used to develop such a model, but 30%-87% of mice die when administered an intraperitoneal injection of 40 mg kg-1. In the present study, hematoxylin and eosin staining, Western blot, immunofluorescence, and quantitative real-time polymerase chain reaction were used to test the effects of busulfan exposure in a mouse model that received two intraperitoneal injections of busulfan at a 3-h interval at different doses (20, 30, and 40 mg kg-1) on day 36 or a dose of 40 mg kg-1 at different time points (0, 9, 18, 27, 36, and 63 days). The survival rate of the mice was 100%. When the mice were treated with 40 mg kg-1 busulfan, dramatic SSC depletion occurred 18 days later and all of the germ cells were cleared by day 36. In addition, the gene expressions of glial cell line-derived neurotrophic factor (GDNF), fibroblast growth factor 2 (FGF2), chemokine (C-X-C Motif) ligand 12 (CXCL12), and colony-stimulating factor 1 (CSF1) were moderately increased by day 36. A 63-day, long-term observation showed the rare restoration of endogenous germ cells in the testes, suggesting that the potential period for SSC transplantation was between day 36 and day 63. Our results demonstrate that the administration of two intraperitoneal injections of busulfan (40 mg kg-1 in total) at a 3-h interval to mice provided a nonlethal and efficient method for recipient preparation in SSC transplantation and could improve treatments for infertility and the understanding of chemotherapy-induced gonadotoxicity.