细胞工程资料.docVIP

1、Why cultivate animal cells in the 5 % CO2 incubator? What Particular care should be taken during the experiment? （1）The cells are grown in an atmosphere of 5% CO2 because the medium used is buffered with sodium bicarbonate and the pH must be strictly maintained. （2）Culture flasks should have loosened caps to allow for sufficient gas exchange. Cells should be left out of the incubator for as little time as possible and the incubator doors should not be opened for very long. The humidity must also be maintained for those cells growing in tissue culture dishes, so a pan of water is kept filled at all times. 2、What laboratory tests are used to identify human embryonic stem cells? （1）growing and subculturing the stem cells for many months. This ensures that the cells are capable of long-term self-renewal. （2）using specific techniques to determine the presence of surface markers that are found only on undifferentiated cells. Another important test is for the presence of a protein called Oct-4, which undifferentiated cells typically make. （3）examining the chromosomes under a microscope. （4）determining whether the cells can be subcultured after freezing, thawing, and replating. （5）testing whether the human embryonic stem cells are pluripotent by (a) allowing the cells to differentiate spontaneously in cell culture;(b) manipulating the cells so they will differentiate to form specific cell types; or (c) injecting the cells into an immunosuppressed mouse to test for the formation of a benign tumor called a teratoma. 3、What is transgenic animal? How to produce transgenic mice by the embryonic stem cell method? A transgenic animal is one that carries a foreign gene that has been deliberately inserted into its genome. The embryonic stem cell method involves: （1）Make your DNA. （2）Transform ES cells in culture. （3）Select for successfully transformed cells. （4）Inject these cells into the inner cell mass (ICM) of mouse blastocysts. （5）Embryo transfer. （6）Test her offspring