Fatima Efendic

Mouse embryonic stem cells (mESCs) are pluripotent stem cells derived from the inner cell mass of the blastocyst, an early stage embryo1,2.

EBs recapitulate many aspects of cell differentiation during early embryogenesis, and play an important role in the differentiation of ES cells into a variety of cell types in vitro

Mouse embryonic stem cells (mESCs) are pluripotent stem cells derived from the inner cell mass of the blastocyst, an early stage embryo1,2.

They are pluripotent in that they are able to differentiate into all derivatives of the primary germ layers, including ectoderm, endoderm, and mesoderm, thus generating every cell type in the body. mESCs also contribute to a wide range of adult tissues in chimeras with intact embryos, including the germ cells.

mEB formation methode:

Materials Needed

mESCs growing on MEF feeder culture

Mouse Embryonic Stem Cell (mESC) EB Medium

(mESC) EB Medium with 10% FBS

scalper,santrifuger,petri dishes,pipets.

Preparing AF-Coated Dishes

Cover the whole surface of a 100-mm tissue culture-treated dish with Attachment Factor (AF) solution and incubate the vessels for 30 minutes at 37°C or for 24 hours at room temperature.

Passaging mESCs into EbsPrepare EB medium on 37°C water bathFrom new dish asparate gelatin From petri with mESCs remove old mediumAdd 2 ml EB medium With scraper unstik cells(do gently)Gentle pipetting,put cells into new petri dish(if there is more than one dish with mESCc use same proces)Comlite medium on 10 ml.

Every two day changing medium using EB medium

After 1 weeks changingdish and use MEF

Every 2 days checking results

After 3 weeks its possible to see cell beating

mouse embryoid body beating cardiomyocytes