2. Mouse embryonic stem cells (mESCs) are
pluripotent stem cells derived from the
inner cell mass of the blastocyst, an early
stage embryo1,2.
3. 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.
4. 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.
5. 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.
6. 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.
7.
8. Passaging mESCs into Ebs
Prepare EB medium on 37°C water bath
From new dish asparate gelatin
From petri with mESCs remove old medium
Add 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.
9. 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
