The Method of Rodent Whole Embryo Culture using the Rotator-type Bottle Culture System

In developmental Biology, there are various animals that are used for the experiment. Hor agriculture technique of rot embryos has been established in 1950s and developed before establishment of modern genetic manipulation in Hawaiian agriculture. Weakened director directly target areas of loving with die or transfect of genes since embryos can be manipulated and as a microscope, therefore whole embr culture is very useful tool.

When we want to study the dynamic development process of maorian embryos after post implantation development. Growth of maorian embryos are critically dependent on the functional presenter, which limits the culture period. In general, we can culture embryos during the period of embryonic day 8.5 to 14.5 in the rat, and E 6.5 and 12.5 in the mouse.

In the laboratory, we culture rat and mouse embryos using a rotating culture system. This culture system is most popular and suitable for culture of rat and mouse embryos during lead gestation. That is after E 11.5 and 9.5 in red and mice.

In this video, we would like to introduce our standard protocol for hungry culture using E 12.5 RU embryos. In this condition, the embryo can be cultured for two days, which is well enough to examine the Various effective manipulation in hor agriculture. There are Two critical steps for the success.

First, the dissection procedure should be accurate not to damage the embryo, especially rod vessels. Second, the procedure should be as quick as possible. This is especially true for the older embryos.

For example, in case of the E 10.5 mouse embryos, that it's they should be transferred within 30 minutes. We applied whole embryo culture technique for pack six mutant right embryos. For example, we analyze migration pattern of neurore cells in the mutant by loving small number of cells with dye or by transplanting level cells in the embryos of white type and muted background.

Furthermore, we have established a method of gene transferring into cultured mouse and red embryos by electro operation that is using electrical shock. We can simply introduce constructs of exogenous genes, dominant negative forms, and siRNAs in the area of the developing brains. Therefore, we can easily analyze gene functions by loss of function and gain function experiments.

We also investigate behavior of cells leveled with GFP or RFP using organ culture or slice culture following, or agriculture. So you can do variety of experiments using Lio culture in combination of other techniques.