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Making Babies in Space May Be Harder Than It Sounds
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The bullet-shaped probe is said to be housing a total of 19 experiments involving microgravity fluid physics, microgravity combustion, space material, space radiation effects, microgravity biological effects and space bio-technology. These include one studying how space radiation affects the genetic stability of fruit flies and rat cells, and a combustion experiment which will test how materials used in spacecraft burn in space to find ways of making safer capsules for future manned missions.
Photo provided by the Institute of Zoology, Chinese Academy of Sciences, shows the blastocysts developed from cells 80 hours after the mouse embryos were launched into space by the satellite, SJ An experiment being run in partnership between the National Space Science Center under the CAS and the European Space Agency will investigate the behavior of crude oil under high pressure, and also on board is equipment to test coal combustion and pollutant formation under microgravity.
The former experiment is aimed at improving scientists' understanding of oil reservoirs buried deep underground, while the latter is expected to help enhance energy efficiency and cut emissions on Earth.
For the First Time Ever, Scientists Have Grown Mammalian Embryos in Space
In a separate development, retrievers for the SJ satellite on Sunday said they will deploy four helicopters to aid ground vehicles in the search for the satellite after its landing next week. Therefore, 3D clinostat experiments will help in predicting and planning real space experiments, which can reduce costs and effort. Thus, a 3D clinostat might have clinical applicability in regenerative medicine to help proliferate stem cells for patients.
However, if this effect is also true for undifferentiated early embryos, they might fail to differentiate or develop to offspring. To evaluate the embryos, some were examined for quality and cell numbers by immunostaining, and others were transferred into pseudopregnant recipient females to test their potential for development to full term. At 6 h after IVF, the clinostat was stopped, and zygotes were collected from the culture flasks. Control studies at 1G were performed using the same batch of preincubated spermatozoa and the same incubator but outside the clinostat Fig.
We also confirmed the normality of zygotes by pronuclear PN staining Fig.
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Similar phenomena were observed in actual space experiments using sea urchin and bull spermatozoa  , . The arrow indicates a polyspermic 3 PN fertilized zygote, and the arrowhead indicates a second polar body. Two to three months later, these offspring grew to adulthood, and randomly selected mice were proven fertile in natural mating f. Therefore, we could not determine the exact fertilization or embryo development rates in each flask.
However, control 1G experiments were performed under exactly the same conditions Fig. It should be noted that these control results are poorer than with conventional IVF methodology  —  ,  — . As discussed above, the use of a flask and a large volume of medium were essential in using the 3D clinostat system, and our preliminary experiments showed that these had negative effects on embryo development. Therefore, we believe that our results are valid. All pairs delivered litters a few months later Fig.
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In mice, the transcription factor Oct4 is expressed throughout preimplantation development and is restricted to the inner cell mass ICM at the blastocyst stage  — . Trophectoderm TE differentiation begins before the downregulation of Oct4 in the outer cells and is marked by Cdx2 expression. Therefore, only Cdx2-positive cells were judged as differentiated, and only Oct4 positive cells were judged as undifferentiated. Nonreactive cells that were DAPI positive nuclear staining were judged to be in metaphase. 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.
Metaphase cells DAPI staining only were counted on triple-merged images e and j. As we acquired 51 focal planes in the z -axis, we could determine the 3D structure of all embryos. In addition to cell number, we also observed the localization of the ICM in blastocysts using a 3D imaging system Fig.
Kojima et al. Possibly this reflects differences between the 1D and 3D clinostat environments. All these offspring appeared normal, and randomly selected animals were later proven fertile by natural mating. However, obviously the recipient females could not be kept in the clinostat, so implantation and gestation in vivo was at 1G. There are two possible explanations.
However, it is unclear whether this also affects mammalian embryos. In other words, the polarization of preimplantation embryos was independent of gravity in this model.
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Unlike studies on early development, far more space research has focused on the latter half of pregnancy, during which many of the early, sensitive phases of development including placentation and early organogenesis normally occur at 1G. Thus far, female rats at several different stages of pregnancy have been launched into space.
These results were consistent with those for other species, especially urodeles  and fish  , . For example, salamander and Medaka fish eggs can be fertilized and develop normally during orbital flight  , .
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ICR CD-1 strain mice were used as pseudopregnant recipient. Hamamatsu, Japan. Fourteen hours after the hCG injection, mice were killed to collect oocytes. The protocols for animal handling and treatment were reviewed and approved by the Animal Care and Use Committee at the same institution. On the other hand, the 3D clinostat system required us to use In this situation, the culture volume is times greater than with the conventional culture system.
Thus, before starting experiments, we had examined the effects of medium volume, of different media, and prolonged culture in a flask with continuous exposure to spermatozoa in terms of embryo development. In terms of extended culture, we have compared the rate of blastocyst development in the flask culture method with the standard protocol outlined above. Although a low embryo-to-culture volume ratio is critical for the optimal development of embryos in vitro  —  it would be impossible to produce sufficient numbers of embryos to compensate for the fold greater volume of flasks compared with microdroplets in dishes.
This suggests that a decrease in the rate of blastocyst development is inevitable when using a relatively large flask. However, we noticed that when embryos were cultured in flasks, blastocyst development was retarded and the embryos required 5 days to develop to the expanded stage.
In addition, it was known that the developmental potential of offspring was unaltered when embryos were cultured in vitro for 4 or 5 days. However, when embryos were cultured for 6 days, the implantation rates of transferred embryos decreased significantly  — . Therefore, in this study, 2-cell embryos were transferred into 1-day-delayed recipient females as usual. However, day 5 blastocysts were transferred into 2-day-delayed day 3 recipient females. Next day, epididymides were collected from two euthanized male mice. A dense sperm mass was squeezed out from the cut epididymis and placed into the flask.
TYH medium was added to fill the flask without any bubbles. The flask was capped and placed in the 3D clinostat, which was then started. During sperm preincubation, oocyte cumulus complex COCs were collected from the oviducts of two or three females, and transferred into a Falcon flask filled with CZB medium that had been equilibrated for one day before use and then placed in the incubator until starting the experiment.
Because COCs were used to enable high fertilization rates, we could not count the number of oocytes in each flask.taylor.evolt.org/pymuv-dosrius-dating-english.php
Mammalian embryos develop in space for the first time
However, after the experiments, we found that each flask had 60— oocytes. After finish the sperm preincubation, the flasks were removed from the 3D clinostat, and 5 ml aliquots of sperm suspension were introduced into the flasks containing the oocytes. CZB medium was added to fill the flask without any bubbles. The 3D clinostat was then started immediately. It took less than 20 min from collecting the sperm suspension to restart the 3D clinostat for IVF. In control experiments, IVF was performed under exactly the same conditions without using the 3D clinostat, and gametes were cultured in the same incubator at the same time Fig.
At 6 h after IVF, all oocytes were collected from the flasks and the fertilization rates were estimated, based on second polar body extrusion. This staining protocol is specific for the female PN. All the embryos were observed using a confocal scanning laser microscope FV, Olympus, Tokyo, Japan. At 24 h or 96 h after IVF, all embryos or oocytes were collected from the flasks.
The rates of 2-cell or blastocyst development were calculated from all collected oocytes, including abnormally ovulated ones these cannot be distinguished from unfertilized oocytes at this stage. Two-cell stage embryos were transferred to oviduct of pseudopregnant ICR mice at 0. These had been mated with a vasectomized ICR male the night before transfer.