The Applications of Alginate Microcapsules in Sheep Spermatogonial Stem Cells Cryopreservation

Abstract: Cryopreservation of animal cells is an essential part of cell biology. In order to improve efficacy of sheep spermatogonial stem cells (SSCs) cryopreservation， alginate microcapsules were used to cryopreserve the sheep SSCs in the present study. Cell viability and culture were evaluated and compared; The products of five SSCspecific proteins， including CDH1， PLZF， GFRα1， OCT-4 and Thy-1 were examined by immunofluorescence after stored in liquid nitrogen for 10 d. The results indicated that alginate microcapsules could increase sheep SSCs viability to 78.7%±1.3%， while stored with single cells suspension was 58.4%±1.4% after stored in liquid nitrogen for 1 d. The viability of sheep SSCs with alginate microcapsules was 78.0%±1.5%， while stored with single cells suspension was 48.1%±0.8% after stored in liquid nitrogen for 10 d. After four days culture， the SSCs with alginate microcapsules embedded can form a typical SSCs clusters after stored in liquid nitrogen for 10 d. Immunofluorescence showed that CDH1， PLZF， GFRα1， OCT-4， Thy-1 were expressed after cryopreservation. Taken together，alginate microcapsules significantly improved the efficacy of sheep SSCs cryopreservation， and maintained the ex pression of sheep SSC-specific genes after thawing. This study indicates a new method for the cryopreservation ofanimal resources by using the alginate microcapsule， especially for the livestock and those endangered species.

CSTR: 32200.14.cjcb.2013.11.0007