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KMID : 1377020140110050379
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Tissue Engineering and Regenerative Medicine
2014 Volume.11 No. 5 p.379 ~ p.386
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Proliferated Leydig cells for engineered testis-like tissue regeneration with testosterone-secreting ability
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Hongda Bi
Xiaoyun Wang
Xin Xing
Wei Liu
Yilin Cao
Guangdong Zhou
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Abstract
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Tissue engineering approach provides a hopeful strategy for reconstructing testis testosterone-secreting functions. However, limited source and low proliferative activity in vitro of Leydig cells (LCs, the main testosteroneproducing cells) makes testis-like tissue regeneration difficult to be achieved. This study explored the feasibility of in vitro expanding LCs and their potential application in testis-like tissue regeneration. LC lineage cells were isolated from Sprague-Dawley (SD) rats by differential adhesion method and cell composition was identified by expressions of 3β-HSD, LHR, LIFR, and c-kit. A modified expansion medium (EM) system was used to test the feasibility of in vitro expanding LC lineage. The results showed that the attached cells reached a high purification of LC lineage (>90%, indicated by positive expression of 3β-HSD) and that EM significantly enhanced proliferation of LC lineage compared to regular medium, which was testified to be related to the presence of stem LCs that was implied by positive expressions of LIFR and c-kit as well as the transition of 3β-HSD expression from negative to positive in partial cells. Importantly, the proliferated LCs showed relatively sustained testosterone-secreting ability in vitro and these cells combined with biodegradable scaffolds successfully regenerated testis-like tissue with sustained testosteronesecreting function in vivo, which was supported by the enhanced serum testosterone level in castrated rats. All these results indicated that the differential adhesion method could efficiently isolate and purify LC lineage and that EM system could efficiently promote proliferation and functional maintenance of LC lineage, providing a good cell source for testes-like tissue regeneration.
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KEYWORD
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leydig cells, proliferation, tissue engineering, testis, testosterone
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