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. 1999 Nov;10(11):3633–3642. doi: 10.1091/mbc.10.11.3633

Figure 7.

Figure 7

Expansion of CAFCs. Lin−Sca-1+c-kit+ cells were purified from BM of the chimeric receptor transgenic mice by FACS sorting. Without preincubation, 100 Lin−Sca-1+c-kit+ cells were incubated on an OP9 stromal cell layer without adding cytokines (░⃞). On the other hand, 100 Lin−Sca-1+c-kit+ cells were incubated with SCF (□), hGM-CSF and SCF (▪), IL-6 and SCF (▧), or IL-11 and SCF (▨) in serum-free suspension culture for 4 d and then washed well with α-medium and incubated on an OP9 stromal cell layer without addition of cytokines. (A) On day 7 of coculture, numbers of cobblestone area were counted. (B) To evaluate the hematopoietic capacity of the cobblestone area, on days 12, 20, and 28, clonogenic cells produced from the cobblestone area during the coculture were analyzed by in vitro colony assay. Assays were performed in triplicate cultures and repeated in two independent experiments.