Two modified density gradient centrifugation methods facilitate the isolation of mouse Leydig cells

Preparation of sufficient mouse Leydig cells (LCs) with high purity is a prerequisite for investigations of the biological/pathological functions of LCs in mouse models. Density gradient centrifugation based on discontinuous Percoll gradients is an effective method (defined as regular method) for LC...

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Published in:Preparative biochemistry & biotechnology 2023-01, Vol.53 (1), p.76-80
Main Authors: Jiang, Jiayang, Zhou, Xiaoman, Gao, Chunliu, Ke, Rongqin, Guo, Qiwei
Format: Article
Language:English
Subjects:
Animal models
Animals
Cell Separation - methods
Centrifugation
Centrifugation, Density Gradient - methods
Density
Density gradient centrifugation
Density gradients
Fluorescence
Interstitial cells
Leydig cell isolation
Leydig Cells
Male
Methods
Mice
Percoll gradient
Purity
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Summary:Preparation of sufficient mouse Leydig cells (LCs) with high purity is a prerequisite for investigations of the biological/pathological functions of LCs in mouse models. Density gradient centrifugation based on discontinuous Percoll gradients is an effective method (defined as regular method) for LC isolation. In this study, we developed two modified methods for LC isolation and compared their performance with that of the regular method. Modified method 1 integrated the crude LCs into the 50% Percoll solution before centrifugation. Modified method 2 sequentially used 50 and 60% Percoll solutions to isolate LCs. The purity of LCs was approximately 88.4, 91.3, and 79.7% derived from the regular, modified 1, and modified 2 methods, respectively. The yields of LCs in the same respective order were approximately 1.7 × 10 5 , 3.9 × 10 5 , and 11.9 × 10 5 cells per 10 8 interstitial cells input. Modified method 1 attained higher purity and yields than those of the regular method. Although the purity of LCs was relatively low for modified method 2, it could be used before further purification by, for example, fluorescence-activated or magnetic-activated cell sorting, owing to its simplicity and high yields. Therefore, our study provided alternative methods to facilitate LC isolation in mice.
ISSN:1082-6068
1532-2297
DOI:10.1080/10826068.2022.2039942