| Budget Amount *help |
¥5,200,000 (Direct Cost: ¥4,000,000、Indirect Cost: ¥1,200,000)
Fiscal Year 2014: ¥1,560,000 (Direct Cost: ¥1,200,000、Indirect Cost: ¥360,000)
Fiscal Year 2013: ¥1,560,000 (Direct Cost: ¥1,200,000、Indirect Cost: ¥360,000)
Fiscal Year 2012: ¥2,080,000 (Direct Cost: ¥1,600,000、Indirect Cost: ¥480,000)
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| Outline of Final Research Achievements |
We crossed transgenic mice having the SPC promoter-driven reverse tetracycline transactivator with transgenic mice expressing histone H2B-GFP protein controlled by a tetracycline responsive regulatory element. Doxycycline was administered to the double transgenic mice from embryonic day 6.5 to postnatal day 14 for labeling. Lungs were harvested at postnatal day 56, and single lung cell suspensions were prepared. LRCs were identified by flow cytometry. Analyses for expression levels of previously reported lung epithelial stem cell markers (Sca-1, CD34, CD90, c-kit, CD24, alpha6-integrin, beta4-integrin) revealed that LRCs were enriched in beta4-integrin, but not in other markers. Both LRCs and H2B-GFPlow cells were also isolated by a fluorescence-activated cell sorter for colony formation assays. LRCs had a higher for epithelial colony formation compared to H2B-GFPlow cells. Fluorescence confocal microscopic analyses revealed that LRCs were located in alveolar area.
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