Immature Testicular Tissue Engineering for Male Fertility Preservation---A Transgenic Mouse Model

Functional male reproductive cells production dependent of germline stem cells (Spermatogonial stem cells-SSCs), after male birth, reproductive cells will form the sperm and pass the genetic message to the next generation.Cryopreservation is a forward-looking medical applications, especially underage cancer patients and endangered species. However, different frozen protection agents have different cell biological toxicity and protective. Cryopreservation all cell types of size, complexity, and membrane permeability of testicular tissue brought challenges. Cell scaffold (Scaffold) is very popular a regenerative medicine research, can improve cell viability and growth rate. With bioluminescence allograft can clearly identify the reproductive growth after cell transplantation, synchronize conduct iso-transplantation and in vivo tracking. The objective of this study is to applications cell scaffold isograft to enhance the spermatogonial stem cells after cryopreservation in order to facilitate research and clinical applications. 1. Never have the reproductive ability of newborn mice achieved testicular tissue, followed by cryopreservation or with Scaffold assessment Scaffold transplanted to the scrotum in the reproductive ability of immature orchiectomized mice in vivo repair and regeneration of fresh the testes and frozen testicular organization circumstances. 2. Scaffold bracket when the tissue regeneration template is the basis of regenerative medicine. Cells of different materials have different reactions. Chemistry, mechanics, and structure of the material has a strong influence regardless of cell attachment, proliferation, migration or differentiation. Scaffolds for three-dimensional culture, cells often have different performance. However, current knowledge of the biological material is still phenomenology. The study the material in order to make more progress of tissue engineering, stem cell differentiation is in need. 3. Application of non-invasive bioluminescence could test the number of cell survival and proliferation by MTT assay and BrDU assay to assess the survival and proliferation rate of testicular tissue, accompanied by a common cell activity analysis to assess the reproductive ability of cryopreservation forming role of repair the SSCs for mice reconstruction spermatogenic effect. 4. Fertility assay after immature testicular engineering is applied to evaluate the efficiency and the impact on male offspring (F1 to F3).