(K) Morphology of primed state iNANOG cells (without Dox) and Dox induced iNANOG cells (passage 9, P9) in the 2iL medium.
Cells passage was performed every 3 4 days and suspensions of each cell line were produced from confluent cultures using 10× TrypLE™ (gibco® by life technologiesTM, USA).
Culture of human breast adenocarcinoma cell line MCF-7 cells (passage 30 35, American Type Culture Collection) and their uptake of the coumarin-6-loaded nanoparticles were performed in the same way.
Primary cultures were passed to disperse the colony-forming cells (passage 1).
Undifferentiated ES cells (passage 25) were transfected with Lipofectamine 2000 (Invitrogen) according to the manufacturer instructions.
The only exception was the 8743THA isolate that was adapted on BSR cells (passage 22).
PC-3 and LNCaP human prostate cancer cells (passage ranges 32 38 and 14 19, respectively), and R3327 AT-1 rat prostate cancer cells (passage range 49 59) were available at the Department.
The expression levels of GATA4 inversely correlate with passage number of the cells: HOSE.96 (or C.96) cells, passage #3, have high GATA4 expression; HOSE.47 cells, passage #5, have a lower GATA4 level; and HOSE.63 cells, passage # 12, have a much reduced expression level (Figure 5B).
Importantly, only cultures containing E/M cells were able to form tumors in SCID-beige mice within 4 months when 105 cells (passage 6) were injected (Figure 1D).
Next we evaluated the expression of SMC-specific genes in the hESC-derived cells (passage 4) by quantitative RT-PCR (Figure 2).
The cells (passage number 5-8) in 35-mm or 100-mm dishes or glass coverslips were used for all the experiments.
