Briefly, cross-linking between DNA and protein was preformed by adding formaldehyde (Merck) directly into the culture medium of mouse embryonic fibroblasts (MEF) derived from wt and Hpse-KO mice.
5 × 103 T241 fibrosarcoma cells or PancO2 cells were seeded in 96-well plates and serum-starved for 6 hours before adding conditioned medium of murine embryonic fibroblasts grown in DMEM containing 10% FCS or stimulated with serum-free DMEM containing 3 ng/mL recombinant human TGF-β1 for 24 hours.
Recombinant human kallistatin was secreted into the serum-free medium of cultured human embryonic kidney cells (HEK293T), and culture medium was concentrated by ammonium sulfate precipitation followed by nickel-affinity and heparin-affinity chromatography as previously described [ 25, 26].
We used two assays of myogenic regenerative activity: the proliferation of human and mouse myoblasts by incorporation of BrdU, and their resistance to differentiation in this otherwise low-mitogen differentiation medium, as measured by expression of embryonic myosin heavy chain (eMyHC).
Even during his childhood, in the Woodhaven section of Queens in the 1940s and '50s, Mr. Turner was captivated by the power and possibilities of the embryonic medium of television.
It should be noted that interaction of MPO with platelets did not depend on the presence in the medium of donors' plasma components, embryonic calf serum or such sugars as α-methyl-D-mannoside, lactose or N-acetyl-D-glucosamine at the final concentration of 60 mM (data not shown).
The addition of exogenous FGF4 and heparin to culture medium allows the derivation of murine multipotent TS cells when cells are grown in the presence of embryonic fibroblast conditioned medium, or on feeders [4], [12].
For example, when cultured in suspension, ES cells in identical medium form embryonic bodies, which consist of cells of the three germ layers.
The hourglass effect refers to higher morphological divergence at earlier or later stages of embryonic developmental, compared to medium stages.
Primary neurons were obtained from the cerebral cortex of embryonic day 17 (E17)/E18 rat embryos and cultured in the Neurobasal medium as reported previously.
