Establishment Of Adenine-induced Recipient Model For Transplantation Of Mouse Spermatagonial Stem Cells

Spermatogonial stem cells (SSCs) are adult stem cells which have the ability toself-renewal and differentiate to spermatogenic cells. After transplanted to recipient testes,SSCs have the potential to differentiate. Preparation of recipients is essential for successfultransplantation. Adenine is widely used to treat cancers, and is also used in the establishmentof chronic renal failure model. Recently, researchers found that adenine could reduce thenumber of sperm.The rats and mice were treated with adenine in order to establish the male sterilityanimal model. We observed the weight, testis index, epididymis index and histologicalchanges of testes. SSCs transplantation was performed to authenticate the recepients. Themain findings are as follows:1.15of2-3months male rats were randomly divided into three groups, the control group,150mg/kg and300mg/kg adenine-treated group. After three weeks, the testes of the twoadenine-treated groups were smaller. The300mg/kg group changed significantly, comparedwith the control group. Most of the seminiferous tubules had sperm in150mg/kg group, inspite of the decrease of interstitial cells. Almost all of the seminiferous tubules were absenceof germ cells in300mg/kg group, suggested that300mg/kg dosage could effectively inducemale sterility. The SSCs were labeled with EGFP green fluorescence by lentiviral transduction.Sixty days after transplantation, spermatogenic cells with green fluorescence were observed inthe recipient testes, indicating that the rats treated with adenine could be recipients of mouseSSCs.2.75of4-6weeks male mice were randomly divided into three groups, the control group,300mg/kg and450mg/kg adenine-treated groups. After20days, the testes of twoadenine-treated groups were smaller, compared with the control group. Almost all of theseminiferous tubules were absence of germ cells in the two adenine-treated groups, a greatmany of seminiferous tubules in450mg/kg group had only one layer of cells. The percentageof seminiferous tubules without spermatogenesis was71%and90%in300mg/kg and450mg/kg group, respectively. SSCs were obtained from EGFP transgenic mice and transplanted to300mg/kg group testes. We found the donor-derived spermatogenic cells in theseminiferous tubules of recipients, suggested that male infertile mice could be induced byadenine and may be used as transplant recipients.