For direct generation of colonies, transiently transfected cell cultures were transferred into CHO-A culture medium (Invitrogen) lacking hypoxanthine and thymidine (HT), and seeded at 5000 cells/well in the culture plates.
By eliminating each TF individually from the pool, taking into account the redundancy among TFs, and reiteration, we established that a minimal set of five TFs (ERG, GATA2, LMO2, RUNX1c, and SCL) was able to robustly and reproducibly induce the generation of colonies of round cells in both adult and embryonic fibroblasts.
It is a visceral, powerful, visually ravishing film, occasionally mawkish, sometimes less than plausible – but it's hard not to be moved by the picture it paints of an entire generation of colony boys wiped out before their time.
Induced HOXB4 expression during EB formation did not affect the generation of colony forming cells and repopulating cells in the OP9 and iHOXB4 system or the appearance of surface markers in EB cells.
However, while the default transmission route of Pseudonocardia is vertical, both across generations of colonies and among workers within colonies [ 17, 33], other actinomycetes than Pseudonocardia have also been isolated, both from Acromyrmex and other attine ants, suggesting that additional environmental acquisition is common [ 29, 31, 32, 39, 40].
Overall, the analysis presented here provides the foundation for beginning to evaluate mass reared flies at a genomic level, and to develop tools for better monitoring the quality of these flies over generations of colony production.
After confirming that Cited2Δ/Δ Trp53+/−, Cited2Δ/Δ Trp53−/−, and Cited2Δ/Δ Ink4a/Arf +/− cells from primary colonies lacked Cited2 expression, we demonstrated efficient generation of secondary colonies (data not shown).
Furthermore, the addition of collagen I to fibrin hydrogels promotes cardiac differentiation and increases the generation of contractile colonies (Kong et al., 2013).
Again, REF was significantly better than MEF for generation of reprogrammed colonies by 3 factors.
Queens and workers were observed throughout the winter, suggesting a second generation of bee colonies active during the winter months.
To further examine the role of myosin II in traction generation of the colonies, we cultured mESCs on 8 kPa substrates with blebbistatin (10 µM) for 5 days.
