To propagate spheres in vitro, spheres were collected by gentle centrifugation, dissociated to single cells, and cultured to generate the next generation of spheres [ 7].
Samples from each plate were separately cultured to generate the F1 generation, which was similarly maintained at either 25 mM NaCl or 305 mM NaCl and sexed when individuals reached adulthood.
Both freshly isolated and 2D cultured HMECs generated TDLU-like structures with similar morphologies, suggesting that short-term 2D culture did not significantly change cell behavior (Fig. 3A,D).
SCC cells isolated from tumors can either be cultured to generate a cell line or injected directly into mice.
After twenty-four hours, non-adherent cells were removed and the remaining cells were cultured to generate macrophages.
56 mice were transplanted with either freshly procured DCIS duct segments, propagated xenograft tumors, or organoids and/or human primary cultured cells generated from the DCIS tissue (Figure 1).
Stage 4 R explants cultured alone generated Sox1+ and Pax6+ neural cells of forebrain character, but no Snail2+, HNK-1+ or Ker+ cells (Fig. 5A), whereas Wnt3A blocked the generation of neural cells, and surprisingly induced Snail2+ and HNK-1+ neural crest cells (Fig. 5B).
Accordingly, stage 2 M explants cultured alone generated FoxG1+ cells, but no Rax2+ cells (Fig. 2B).
KW constructed plasmids, inserted NLS by mutagenesis, cultured cells, generated stable cell lines.
Consistent with this idea, transfection of in-vitro-transcribed RNA into cultured cells generates an innate immune response.
