In vitro radiation sensitivity of mouse lung fibroblasts isolated by flow cytometry

Abstract

Recently, we have isolated two major fibroblast cells (Thy-1+, Thy-1-) from mouse LAF1 lung tissue using the anti-Thy-1 antibody expression and fluorescence activated cell sorter. To examine the possibility that x- or gamma-ray-induced pulmonary fibrosis at the late stage of injury could arise from radioresistant cell subpopulations, the radiation sensitivities of Thy-1+ and Thy-1- cells were evaluated by the colony forming assay. Cell survival curves, repair of potentially lethal damage (PLD) and sublethal damage (SLD), and cell-age response curves were obtained after Cs-137 gamma-ray irradiation. The cell survival curves measured after 0-10 Gy gamma-ray showed that Thy-1+ cells were slightly more radioresistant than Thy-1- cells. The D0, n, alpha, and beta values measured from the survival curves also confirmed this observation. After a single dose of 10 Gy, a small amount of PLD repair was observed in Thy-1- cells, while no PLD repair was found in Thy-1+ cells. Although the initial cell survival level of Thy-1- cells was lower, the final survival levels of Thy-1+ and Thy-1- cells became identical at 8 h after irradiation due to the PLD repair. After split-dose irradiation of 4 Gy followed by 4 Gy, a similar extent and rate of SLD repair was found in Thy-1+ and Thy-1- cells. Cell-age response curves were obtained from irradiated G0/G1, S, and G2M cells separated by centrifugal elutriation and irradiated with 8 Gy gamma-ray. The results indicated that Thy-1+ and Thy-1- cells had a similar S resistant, and G1, G2M-sensitive radiation cell-age response curve. This study suggests that the selection of radioresistant lung fibroblast may not be responsible for the development of lung fibrosis in irradiated LAF1 mouse.