Previous reports have shown that human embryonic stem (ES) cells can be maintained in feeder-free condition.
Lastly, Huv-iPS cell lines displayed a normal karyotype (Figure 5D), and have been maintained in feeder-free conditions for over 40 passages.
ESCs were maintained in feeder-free conditions using ESGRO complete (Chemicon) serum-free medium supplemented with 1 5% FBS to enhance proliferation.
We found that mouse ES cells could be maintained in feeder-free and serum-free culture conditions without LIF in simulated microgravity using the 3D-clinostat.
To enhance cell survival, we generated EBs from hESC which were maintained in feeder-free conditions, after which the differentiation medium was replaced with hESC growth medium lacking the mitogen bFGF.
When hESC maintained in feeder-free conditions were cultured in hESC medium lacking bFGF, but with omission of the EB step, they survived well, but SPIE induced minimal DA differentiation (data not shown).
The hESC cell line WA01 (WiCell H1) was maintained in feeder-free conditions.
Human ES cell line H9 (WiCell, Madison, WI) was maintained in feeder-independent conditions, using Synthemax SC-II Substrate (Corning) and grown in TeSR-E8 (Stemcell Technologies, Canada).
IMR90-4 induced pluripotent stem cells (iPSCs), H9 human embryonic stem cells (hESCs), and DF-19-9-11T iPSCs were maintained in feeder-free conditions on Matrigel (BD Biosciences) in mTeSR1 media (WiCell Research Institute).
hES cells lines H9 were maintained in a feeder-free culture over matrigel-coated (BD Biosciences) six-well plate in bFGF-hFLSCs-CM.
These cells were maintained in a feeder-free culture system using mTesR1 Stem Cell Technologiess, Vancouver, BC, Canada) and Matrigel (BD Bioscience, Bedford, MA) with 1% (v/v) penicillin-streptomycin. TALEN proteins, including standard TALENs and TAT-TALENs were prepared for treatment as follows: TALEN proteins were diluted into serum-free high-glucose DMEM.
