Freezing and Thawing of MEFs
|Author: Shalini Jain and Hariom Yadav|
|Affiliation: Animal Biochemistry Division, National Dairy Research Institute, Karnal-132001, Haryana, India|
|Date Added: Mon Feb 02 2009|
|Date Modified: Mon Feb 02 2009|
|Abstract: This protocol is very useful to the researchers working on primary mouse embryonic fibrobasts and gene cell lines|
During experimental needs the preservation of cell lines and their recovery after preservation are important steps in laboratory. Cryopreservation is a best procedure to preserve the cell lines, but accuracy during handling is an important factor which gives high yield of cells after storage. Rapid freezing and slow thawing gives low recovery of cells after cryopreservation, thereby slow freezing gives maximal survival of mammalian cells provided that a cryoprotective agent and rapid thawing are used. It could be supposed, therefore, that this type of freezing would cause minimal structural and functional damage to the cells.
Embryonic stem cells are usually grown on a layer of mitotically inactivated primary mouse embryonic fibroblasts (MEFs) to promote growth and prevent differentiation. These cells stop dividing after a couple of passages, so embryonic fibroblasts need to be isolated freshly from time to time. To avoid isolation at time to time we need to be preserve them safely.
Materials and reagents