In vitro culture of spermatogonial stem cells isolated from adult alpaca (Vicugna pacos) testes analysed with Dolichos biflorus by flow cytometry

Martha Esther Valdivia Cuya, Jhakelin Reyes, Zeze Humberto Bravo Gutierrez, Christian Cancho, Sergio Castañeda, Orlando Limaymanta, Patricia Gloria Woll Toso, Alexei Vicent Santiani Acosta, Gustavo F. Gonzales

Research output: Contribution to journalArticlepeer-review

1 Scopus citations

Abstract

Spermatogonial stem cell (SSC) is known for its self-renewal capacity. We have studied the in vitro proliferation of isolated SSC from adult alpaca (Vicugna pacos) testes. A total of 107 samples were evaluated of which 31 were evaluated at baseline, 36 were cultivated in DMEM and 40 in STEMPRO media. Half of the cultivated samples was analysed after 14 days, and the rest after 21 days. Round cell subpopulations were identified with FITC-DBA by flow cytometry: strongly positive DBA (sDBA+) as SSC, weakly positive DBA (wDBA+) as in early differentiation and negative DBA as differentiated. At the beginning, 4.16% of the cells were SSC, 37.61% wDBA+ while 54.12% were DBA−. After 14 days, 42.28% of SSC, 44.68% wDBA+ and 11.07% DBA− were found in DMEM while 47.09% of SSC, 32.57% wDBA+ and 18.48% DBA− in STEMPRO. After 21 days 38.66% were SSC, 52.78% wDBA and 7.65% DBA− in DMEM and on STEMPRO media 47.92% SSC, 44.20% wDBA+, 4.93% DBA−. There is a significant difference between the number of initial and SSC cultivated, as well as between DBA− (p < 0.05), while there is no significant difference between the wDBA+ (p > 0.05). Our results suggest that both culture media are appropriate for the in vitro proliferation of alpacas SSC.

Original languageEnglish
Article numbere13269
JournalAndrologia
Volume51
Issue number6
DOIs
StatePublished - Jul 2019
Externally publishedYes

Bibliographical note

Funding Information:
This study was supported by Grants N 161001091 and N B17100101

Publisher Copyright:
© 2019 Blackwell Verlag GmbH

Copyright:
Copyright 2019 Elsevier B.V., All rights reserved.

Keywords

  • DBA
  • alpaca
  • flow cytometry
  • in vitro culture
  • spermatogonial stem cell

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