Ultra-Fast and Optimized Method for the Preparation of Rodent Testicular Cells for Flow Cytometric Analysis

Biological Procedures Online

Rosana Rodríguez-Casuriaga¹  Adriana Geisinger²  Beatriz López-Carro³  Valentina Porro⁴  Rodolfo Wettstein¹  Gustavo A Folle³ 

[1] Departamento de Biología Molecular, Instituto de Investigaciones Biológicas Clemente Estable (IIBCE), CP 11600, Avda., 3318, Montevideo, Uruguay

[2] Facultad de Ciencias, Montevideo, Uruguay

[3] Servicio de Citometría de Flujo y Clasificación Celular (SECIF), IIBCE, Montevideo, Uruguay

[4] Unidad de Biología Celular, Instituto Pasteur de Montevideo (IPMONT), Montevideo, Uruguay

关键词: Spermatogenesis;    Gene expression;    Flow cytometry;   

Others  :  797194
DOI  :  10.1186/s12575-009-9003-2

 received in 2009-01-23, accepted in 2009-02-07,  发布年份 2009

【 摘 要 】

Homogeneity of cell populations is a prerequisite for the analysis of biochemical and molecular events during male gamete differentiation. Given the complex organization of the mammalian testicular tissue, various methods have been used to obtain enriched or purified cell populations, including flow cell sorting. Current protocols are usually time-consuming and may imply loss of short-lived RNAs, which is undesirable for expression profiling. We describe an optimized method to speed up the preparation of suitable testicular cell suspensions for cytometric analysis of different spermatogenic stages from rodents. The procedure takes only 15 min including testis dissection, tissue cutting, and processing through the Medimachine System (Becton Dickinson). This method could be a substitute for the more tedious and time-consuming cell preparation techniques currently in use.

【 授权许可】

   
2009 Rodríguez-Casuriaga et al.

【 图 表 】

Figure 1.

Figure 2.

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