Effects of the conditioned medium of mesenchymal stem cells on mouse oocyte activation and development
Mesenchymal stem cells (MSCs) have been reported to secrete a variety of cytokines and growth factors acting as trophic suppliers, but little is known regarding the effects of conditioned medium (CM) of MSCs isolated from femurs and tibias of mouse on the artificial activation of mouse oocytes and o...
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Associação Brasileira de Divulgação Científica
2009-06-01
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doaj-ec57ec619bbf4bca97e1dbe77f5686c62020-11-24T21:02:01ZengAssociação Brasileira de Divulgação CientíficaBrazilian Journal of Medical and Biological Research0100-879X1414-431X2009-06-0142650651410.1590/S0100-879X2009000600006Effects of the conditioned medium of mesenchymal stem cells on mouse oocyte activation and developmentD.Q. FengY. ZhouB. LingT. GaoY.Y. ShiH.M. WeiZ.G. TianMesenchymal stem cells (MSCs) have been reported to secrete a variety of cytokines and growth factors acting as trophic suppliers, but little is known regarding the effects of conditioned medium (CM) of MSCs isolated from femurs and tibias of mouse on the artificial activation of mouse oocytes and on the developmental competence of the parthenotes. In the current study, we investigated the effect of CM on the events of mouse oocyte activation, namely oscillations of cytosolic calcium concentration ([Ca²+]i), meiosis resumption, pronucleus formation, and parthenogenetic development. The surface markers of MSCs were identified with a fluorescence-activated cell sorter. The dynamic changes of the spindle and formation of pronuclei were examined by laser-scanning confocal microscopy. Exposure of cumulus-oocyte complexes to CM for 40 min was optimal for inducing oocyte parthenogenetic activation and evoking [Ca²+]i oscillations similar to those evoked by sperm (95 vs 100%; P > 0.05). Parthenogenetically activated oocytes immediately treated with 7.5 µg/mL cytochalasin B (CB), which inhibited spindle rotation and second polar body extrusion, were mostly diploid (93 vs 6%, P < 0.01) while CB-untreated oocytes were mostly haploid (5 vs 83%, P < 0.01). Consequently, the blastocyst rate was higher in the CB-treated than in the CB-untreated oocytes. There was no significant difference in developmental rate between oocytes activated with CM and 7% ethanol (62 vs 62%, P > 0.05), but the developmental competence of the fertilized oocytes was superior to that of the parthenotes (88 vs 62%, P < 0.05). The present results demonstrate that CM can effectively activate mouse oocytes, as judged by the generation of [Ca²+]i oscillations, completion of meiosis and parthenogenetic development.http://www.scielo.br/scielo.php?script=sci_arttext&pid=S0100-879X2009000600006Mesenchymal stem cellsOocyte activationParthenogenesisCalciumCytochalasin B |
collection |
DOAJ |
language |
English |
format |
Article |
sources |
DOAJ |
author |
D.Q. Feng Y. Zhou B. Ling T. Gao Y.Y. Shi H.M. Wei Z.G. Tian |
spellingShingle |
D.Q. Feng Y. Zhou B. Ling T. Gao Y.Y. Shi H.M. Wei Z.G. Tian Effects of the conditioned medium of mesenchymal stem cells on mouse oocyte activation and development Brazilian Journal of Medical and Biological Research Mesenchymal stem cells Oocyte activation Parthenogenesis Calcium Cytochalasin B |
author_facet |
D.Q. Feng Y. Zhou B. Ling T. Gao Y.Y. Shi H.M. Wei Z.G. Tian |
author_sort |
D.Q. Feng |
title |
Effects of the conditioned medium of mesenchymal stem cells on mouse oocyte activation and development |
title_short |
Effects of the conditioned medium of mesenchymal stem cells on mouse oocyte activation and development |
title_full |
Effects of the conditioned medium of mesenchymal stem cells on mouse oocyte activation and development |
title_fullStr |
Effects of the conditioned medium of mesenchymal stem cells on mouse oocyte activation and development |
title_full_unstemmed |
Effects of the conditioned medium of mesenchymal stem cells on mouse oocyte activation and development |
title_sort |
effects of the conditioned medium of mesenchymal stem cells on mouse oocyte activation and development |
publisher |
Associação Brasileira de Divulgação Científica |
series |
Brazilian Journal of Medical and Biological Research |
issn |
0100-879X 1414-431X |
publishDate |
2009-06-01 |
description |
Mesenchymal stem cells (MSCs) have been reported to secrete a variety of cytokines and growth factors acting as trophic suppliers, but little is known regarding the effects of conditioned medium (CM) of MSCs isolated from femurs and tibias of mouse on the artificial activation of mouse oocytes and on the developmental competence of the parthenotes. In the current study, we investigated the effect of CM on the events of mouse oocyte activation, namely oscillations of cytosolic calcium concentration ([Ca²+]i), meiosis resumption, pronucleus formation, and parthenogenetic development. The surface markers of MSCs were identified with a fluorescence-activated cell sorter. The dynamic changes of the spindle and formation of pronuclei were examined by laser-scanning confocal microscopy. Exposure of cumulus-oocyte complexes to CM for 40 min was optimal for inducing oocyte parthenogenetic activation and evoking [Ca²+]i oscillations similar to those evoked by sperm (95 vs 100%; P > 0.05). Parthenogenetically activated oocytes immediately treated with 7.5 µg/mL cytochalasin B (CB), which inhibited spindle rotation and second polar body extrusion, were mostly diploid (93 vs 6%, P < 0.01) while CB-untreated oocytes were mostly haploid (5 vs 83%, P < 0.01). Consequently, the blastocyst rate was higher in the CB-treated than in the CB-untreated oocytes. There was no significant difference in developmental rate between oocytes activated with CM and 7% ethanol (62 vs 62%, P > 0.05), but the developmental competence of the fertilized oocytes was superior to that of the parthenotes (88 vs 62%, P < 0.05). The present results demonstrate that CM can effectively activate mouse oocytes, as judged by the generation of [Ca²+]i oscillations, completion of meiosis and parthenogenetic development. |
topic |
Mesenchymal stem cells Oocyte activation Parthenogenesis Calcium Cytochalasin B |
url |
http://www.scielo.br/scielo.php?script=sci_arttext&pid=S0100-879X2009000600006 |
work_keys_str_mv |
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