Fig. 5.

BMSC increase proliferation of NHLF cells when grown in coculture. Target NHLF were plated at 1 × 10⁴ cells/well on the bottom surface of Transwell plates (0.3-μm pore size) and cultured in serum-containing fibroblast basal medium (FBM) for 24 h. Subsequently, MSC or NHLF cells were plated at in the top well at 4 × 10⁴ cells/well in serum-containing α-minimum essential medium (AMEM; CBMSC), Iscove's modified Dulbecco's medium (IMDM; BMSC), or FBM (NHLF). Control wells contained AMEM only in the top wells. After 24 h, media in the top wells was replaced with SF AMEM (CBMSC) or SF IMDM (BMSC), and media in the bottom wells was replaced with SF FBM. After 72 h, target NHLF cells were enumerated in trypan blue. It is clear that the MSC in the top well significantly increased target cell numbers in the lower well by means of a diffusible factor. Data represent the means ± SE from triplicate wells for each treatment group. *Significantly different from AMEM and from NHLF, P < 0.05.