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. Author manuscript; available in PMC: 2011 May 1.
Published in final edited form as: Int J Oncol. 2010 May;36(5):1165–1173. doi: 10.3892/ijo_00000599

Figure 5.

Figure 5

To determine the involvement of cell cycle regulatory molecules and the RAS signaling pathway in glioma cells in contact with hUCB stem cells, a modified Boyden’s chamber was used as previously described. The chamber consisted of a porous membrane (0.22 μm pore size), which was coated with Matrigel to facilitate the growth of cells on both the upper and lower surfaces. Human umbilical cord blood stem cells stained with lipophilic cell tracker red dye were grown on the upper surface and glioma cells (SNB19, U87, 5310 or 4910) were grown on the lower surface. The cells were allowed to grow for 72 hrs under standard cell culture conditions, after which the lower glioma surface layer was carefully scraped out and total RNA isolated as per standard protocol. Real time RT-PCR analysis was carried out using primers specific for cell cycle regulatory molecules and the RAS signaling pathway (A). Agarose gels were run to validate real time RT-PCR results, which indicated changes in key cell cycle regulatory and RAS pathway molecules (B).