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(Reference retrieved automatically from Web of Science through information on FAPESP grant and its corresponding number as mentioned in the publication by the authors.)

alpha-6 Integrin Expression in Bovine Spermatogonial Cells Purified by Discontinuous Percoll Density Gradient

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Author(s):
de Barros, F. R. O. [1] ; Worst, R. A. ; Saurin, G. C. P. ; Mendes, C. M. ; Assumpcao, M. E. O. A. ; Visintin, J. A.
Total Authors: 6
Affiliation:
[1] Univ Sao Paulo, Dept Anim Reprod, VRA, Sch Vet Med & Anim Sci, Lab Vitro Fertilizat Cloni, BR-05508270 Sao Paulo - Brazil
Total Affiliations: 1
Document type: Journal article
Source: REPRODUCTION IN DOMESTIC ANIMALS; v. 47, n. 6, p. 887-890, DEC 2012.
Web of Science Citations: 9
Abstract

The study of spermatogonial stem cells (SSCs) provides a model to better understand adult stem cell biology. Besides the biomedical potential to perform studies of infertility in many species, SSCs hold a promising application at animal transgenesis. Because stem cells are thought to be associated with basement membranes, expression of alpha-6 integrin has been investigated as a marker of type A spermatogonial cells, which are considered SSCs because of their undifferentiated status and self-renewal ability. In this manner, the aim of this study was to isolate type A SSCs from adult bulls by a two-step enzymatic procedure followed by a discontinuous Percoll density gradient purification and verify the expression of alpha-6 integrin by flow cytometry and real-time RT-PCR before and after Percoll purification. Spermatogonial cells were successfully obtained using the two-step enzymatic digestion. An average of 1 x 10(5) viable cells per gram of testis was isolated. However, the discontinuous Percoll did not purify isolated cells regarding alpha-6 integrin expression. Flow cytometry analysis demonstrated no differences in the alpha-6 integrin expression between cell samples before and after Percoll purification (p = 0.5636). The same was observed in the real-time PCR analysis (p > 0.05). In addition to alpha-6 integrin, the expression of GFR alpha-1 and PGP9.5, known bovine SSCs markers, was detected in all samples studied. Considering that Percoll can reduce cell viability, it is possible to conclude that Percoll density gradient is not suitable to purify bovine SSC, according to alpha-6 integrin expression. (AU)

FAPESP's process: 10/51654-2 - Use of spermatogonial stem cells to produce transgenic animals
Grantee:José Antonio Visintin
Support Opportunities: Regular Research Grants