|
|
|
|
|
|
|
|
|||||
| Home Help Feedback Subscriptions Archive Search Table of Contents | |||||
|
Fig. 3. Laminin supports long-term proliferation of Dlk+ cells. (A,B) The number of large colonies, containing more than 100 cells, observed at 5, 14 and 21 days after plating Dlk+ cells on type IV collagen (A) and laminin (B). While on type IV collagen, the number of large colonies decreased during long-term culture (A), some medium-sized colonies continued to expand on laminin after 5 days and became large colonies at 14 days (red portion of bar) and at 21 days (yellow portion of bar) (B). Large colonies were marked at the 5th day after plating and their sizes were checked at days 14 and 21. The colonies that contained more than 100 cells on day 14 and 21 were also marked. Data shown are the number of colonies formed from 1000 Dlk+ cells. The culture was repeated independently four times, and average values of colony number are shown. Error bars represent standard deviation for the total number of large colonies at each time point. Student's t-test was performed and P values are shown in A and B. (C) Expression of Ki67 after 21 days of culture. About 20% of the cells on laminin expressed Ki67 (panel 3), whereas most of the cells on type IV collagen did not (panel 2). Cultured cells were dissociated from plates after 21 days of culture, fixed in 4% paraformaldehyde, and permeabilized in methanol. After incubation with mouse IgG (panel 1) or mouse anti-human Ki67 mAb (panels 2 and 3) followed by treatment with FITC-conjugated anti-mouse IgG, the expression of Ki67 was examined by using FACScaliber. (D) The colony derived from a single Dlk+ cell grew continuously on laminin. The same colony was photographed at 5, 14, 21 and 31 days after plating. Bar, 200 µm.
|
