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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Main Authors: D.Q. Feng, Y. Zhou, B. Ling, T. Gao, Y.Y. Shi, H.M. Wei, Z.G. Tian
Format: Article
Language:English
Published: Associação Brasileira de Divulgação Científica 2009-06-01
Series:Brazilian Journal of Medical and Biological Research
Subjects:
Online Access:http://www.scielo.br/scielo.php?script=sci_arttext&pid=S0100-879X2009000600006
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spelling 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
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