An autofluorescence-based isolation of Leydig cells for testosterone deficiency treatment.
An autofluorescence-based isolation of Leydig cells for testosterone deficiency treatment.
复制标题
基于自发荧光的间质细胞分离，用于治疗睾酮缺乏症。
DOI：
10.1016/j.mce.2021.111389
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发表时间：
2021
影响因子：
4.1
通讯作者：
Deng Chunhua
中科院分区：
文献类型：
--
作者：
Luo Peng;Feng Xin;Deng Ronghai;Wang Fulin;Zhang Yadong;Li Xiangping;Zhang Min;Wan Zi;Xiang Andy Peng;Xia Kai;Gao Yong;Deng Chunhua
间质细胞（Leydig cells，LC）的有效纯化方法可以促进功能研究和移植治疗。然而，目前从睾丸中纯化LC的方法仍远不能令人满意。本研究发现，从出生到成年，随着LC的逐渐成熟，睾丸组织中沿着有睾丸自体荧光。通过荧光激活细胞分选（FACS）进一步分离这些自发荧光细胞，并确定其由LC和巨噬细胞组成。为了进一步纯化LC，我们结合了两个荧光通道的流式细胞仪，成功地分离了LC和巨噬细胞。值得注意的是，我们证实所获得的LC不仅具有高纯度、活力和数量，而且具有完整的类固醇生成活性和对促黄体激素的优异反应性。此外，皮下移植分离的LCs可以缓解去势小鼠的睾酮缺乏症状。综上所述，我们建立了一种有效的基于自体荧光的分离LC的方法。这种方法将有助于在未来的成功使用LC的基本和翻译应用。
Effective procedures for the purification of Leydig cells (LCs) can facilitate functional studies and transplantation therapies. However, current methods to purify LCs from testes are still far from satisfactory. Here, we found that testicular autofluorescence existed in the interstitium along with the gradual maturation of LCs from birth to adulthood. These autofluorescent cells were further isolated by fluorescence-activated cell sorting (FACS) and determined to be composed of LCs and macrophages. To further purify LCs, we combined two fluorescence channels of FACS and successfully separated LCs and macrophages. Of note, we confirmed that the obtained LCs not only possessed high purity, viability and quantity but also had intact steroidogenic activity and excellent responsiveness to luteinizing hormone. Moreover, subcutaneous transplantation of isolated LCs could alleviate the symptoms of testosterone deficiency in castrated mice. In summary, we established an effective autofluorescence-based method for isolating LCs. This method will aid in the future success of using LCs for basic and translational applications.