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PLoS One
2009 Aug 25;48:e6753. doi: 10.1371/journal.pone.0006753.
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Detrimental effects of microgravity on mouse preimplantation development in vitro.
Wakayama S
,
Kawahara Y
,
Li C
,
Yamagata K
,
Yuge L
,
Wakayama T
.
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Sustaining life beyond Earth either on space stations or on other planets will require a clear understanding of how the space environment affects key phases of mammalian reproduction. However, because of the difficulty of doing such experiments in mammals, most studies of reproduction in space have been carried out with other taxa, such as sea urchins, fish, amphibians or birds. Here, we studied the possibility of mammalian fertilization and preimplantation development under microgravity (microG) conditions using a three-dimensional (3D) clinostat, which faithfully simulates 10(-3) G using 3D rotation. Fertilization occurred normally in vitro under microG. However, although we obtained 75 healthy offspring from microG-fertilized and -cultured embryos after transfer to recipient females, the birth rate was lower than among the 1G controls. Immunostaining demonstrated that in vitro culture under microG caused slower development and fewer trophectoderm cells than in 1G controls but did not affect polarization of the blastocyst. These results suggest for the first time that fertilization can occur normally under microG environment in a mammal, but normal preimplantation embryo development might require 1G.
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Figure 1. Experimental procedure and 3D clinostat.
Figure 2. Preimplantation development of microgravity (µG)-fertilized and -cultured embryos and development of full-term offspring.Mouse oocytes were fertilized with spermatozoa preincubated under µG. (a) Zygote at 6 h after insemination. (b) Nuclear staining with DAPI. The arrow indicates a polyspermic (3 PN) fertilized zygote, and the arrowhead indicates a second polar body. (c) Two-cell stage embryo cultured for 24 h under µG. (d) Blastocyst cultured for 96 h under µG. (e) Offspring derived from µG-fertilized and -cultured blastocysts. Two to three months later, these offspring grew to adulthood, and randomly selected mice were proven fertile in natural mating (f).
Figure 3. Blastocyst quality after culture under microgravity.The trophectoderm (TE) and inner cell mass (ICM) cell numbers were counted by immunostaining for Cdx2 (a and f, green) or Oct4 (b and g, red), respectively. DNA was stained with DAPI (c and h, blue). Cdx2/Oct4 doubly positive cells were counted on merged images (d and i). Metaphase cells (DAPI staining only) were counted on triple-merged images (e and j). The ICM localization was examined using a three-dimensional (3D) viewer (k and l, and Movie S2 and S3). As we acquired 51 focal planes in the z-axis, we could determine the 3D structure of all embryos. (m) Rates of development to the 2-cell and blastocyst stages in 1G and µG culture systems. (n) Cell numbers and cell types compared between 1G- and µG-cultured blastocysts.
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