Spermatozoa undergo apoptotic changes during the cryopreservation process. These changes, recently termed spermptosis, resemble the cryopreservation induced delayed onset of cell death observed after thawing of somatic cells. Due to its importance in cryobiology, methods to easily identify spermptotic cells are warranted. In this study, a well-validated method for identification of spermatozoa with caspase 3 activity was compared with use of the combination of Hoechst 33342 (H-42) and ethidium homodimer (Eth-1). Live, dead and apoptotic spermatozoa assessed with each method were compared using descriptive statistics and method agreement analysis. No differences were observed in the percentages of spermatozoa in each of the categories investigated with each method. Moreover the method agreement analysis indicated there were consistent findings using both methods The combination H-42/Eth-1 can be successfully used to determine apoptosis in addition to dead and live spermatozoa. Moreover the intensity of H-42 fluorescence (bright and dim populations) allows for distinguishing of live and dead sperm cells.

Gil M.C., Ferrusola C.O., Anel-Lopez L., Ortiz-Rodriguez J.M., Alvarez M., de Paz P., et al. (2018). A simple flow cytometry protocol to determine simultaneously live, dead and apoptotic stallion spermatozoa in fresh and frozen thawed samples. ANIMAL REPRODUCTION SCIENCE, 189, 69-76 [10.1016/j.anireprosci.2017.12.009].

A simple flow cytometry protocol to determine simultaneously live, dead and apoptotic stallion spermatozoa in fresh and frozen thawed samples

Ortiz-Rodriguez J. M.;Pena F. J.
2018

Abstract

Spermatozoa undergo apoptotic changes during the cryopreservation process. These changes, recently termed spermptosis, resemble the cryopreservation induced delayed onset of cell death observed after thawing of somatic cells. Due to its importance in cryobiology, methods to easily identify spermptotic cells are warranted. In this study, a well-validated method for identification of spermatozoa with caspase 3 activity was compared with use of the combination of Hoechst 33342 (H-42) and ethidium homodimer (Eth-1). Live, dead and apoptotic spermatozoa assessed with each method were compared using descriptive statistics and method agreement analysis. No differences were observed in the percentages of spermatozoa in each of the categories investigated with each method. Moreover the method agreement analysis indicated there were consistent findings using both methods The combination H-42/Eth-1 can be successfully used to determine apoptosis in addition to dead and live spermatozoa. Moreover the intensity of H-42 fluorescence (bright and dim populations) allows for distinguishing of live and dead sperm cells.
2018
Gil M.C., Ferrusola C.O., Anel-Lopez L., Ortiz-Rodriguez J.M., Alvarez M., de Paz P., et al. (2018). A simple flow cytometry protocol to determine simultaneously live, dead and apoptotic stallion spermatozoa in fresh and frozen thawed samples. ANIMAL REPRODUCTION SCIENCE, 189, 69-76 [10.1016/j.anireprosci.2017.12.009].
Gil M.C.; Ferrusola C.O.; Anel-Lopez L.; Ortiz-Rodriguez J.M.; Alvarez M.; de Paz P.; Anel L.; Pena F.J.
File in questo prodotto:
Eventuali allegati, non sono esposti

I documenti in IRIS sono protetti da copyright e tutti i diritti sono riservati, salvo diversa indicazione.

Utilizza questo identificativo per citare o creare un link a questo documento: https://hdl.handle.net/11585/927546
 Attenzione

Attenzione! I dati visualizzati non sono stati sottoposti a validazione da parte dell'ateneo

Citazioni
  • ???jsp.display-item.citation.pmc??? ND
  • Scopus 8
  • ???jsp.display-item.citation.isi??? 8
social impact