Premium
Molecular markers of putative spermatogonial stem cells in the domestic cat
Author(s) -
BedfordGuaus SJ,
Kim S,
Mulero L,
Vaquero JM,
Morera C,
AdanMilanès R,
Veiga A,
Raya Á
Publication year - 2017
Publication title -
reproduction in domestic animals
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.546
H-Index - 66
eISSN - 1439-0531
pISSN - 0936-6768
DOI - 10.1111/rda.12819
Subject(s) - biology , stem cell , genetics
Contents Spermatogonial stem cells ( SSC s) are an important tool for fertility preservation and species conservation. The ability to expand SSC s by in vitro culture is a crucial premise for their use in assisted reproduction. Because SSC s represent a small proportion of the germ cells in the adult testis, culture success is aided by pre‐enrichment through sorting techniques based on cell surface‐specific markers. Given the importance of the domestic cat as a model for conservation of endangered wild felids, herein we sought to examine culture conditions as well as molecular markers for cat SSC s. Using a cell culture medium for mouse SSC s supplemented with glial cell‐derived neurotrophic factor ( GDNF ), germ cells from prepuberal cat testes remained viable in culture for up to 43 days. Immunohistochemistry for promyelocytic leukaemia zinc finger ( PLZF ) protein on foetal, prepuberal and adult testis sections revealed a pattern of expression consistent with the labelling of undifferentiated spermatogonia. Fluorescence‐activated cell sorting ( FACS ) with an antibody against epithelial cell adhesion molecule ( EPCAM ) was used to sort live cells. Then, the gene expression profile of EPCAM ‐sorted cells was investigated through RT ‐ qPCR . Notably, EPCAM (+) cells expressed relatively high levels of CKIT ( CD 117 ), a surface protein typically expressed in differentiating germ cells but not SSC s. Conversely, EPCAM (‐) cells expressed relatively high levels of POU domain class 5 transcription factor 1 ( POU 1F5 or OCT 4 ), clearly a germ line stem cell marker. These results suggest that cat SSC s would probably be found within the population of EPCAM (–) cells. Future studies should identify additional surface markers that alone or in combination can be used to further enrich SSC s from cat germ cells.