兔皮肤成纤维细胞在脱细胞真皮支架材料表面生长的研究

Studies on the Growth of Rabbit Skin Fibroblasts on the Surfaces of Acellular Dermal Matrix

为了研究兔皮肤成纤维细胞在脱细胞真皮支架材料表面的生长活性,体外分离、扩增兔皮肤成纤维细胞,鉴定后作为种子细胞接种于脱细胞真皮支架材料上,对细胞在材料上的黏附率及其生长活力进行检测。利用DAPI荧光染色及SEM对细胞在材料上生长情况进行观察,对脱细胞真皮支架材料作为细胞载体的可行性以及用于移植的可行性进行了探讨。结果显示,可分离出单一的成纤维细胞,细胞在材料上的黏附率为96.78%,与在孔板内生长的细胞黏附率无显著性差异(P>0.05);细胞在材料上和在孔板上的生长趋势相似,但是在材料上生长活力较在孔板上小(P<0.01);DAPI荧光染色观察发现,材料上生长的细胞核形完整,且随着培养时间的增加,细胞数量增加;SEM观察结果显示,细胞在材料上能紧密黏附,培养5～10d,细胞在材料上能融合成细胞膜片。提示此脱细胞真皮支架材料可作为成纤维细胞培养载体和移植材料。

The growth of fibroblasts on the acellular dermal matrix （ADM） was studied. The fibroblasts isolated from the skin of an adult New Zealand Rabbit were cultured in vitro and identified subsequently. After the cells were inoculated on the ADM as seeds, the adhesion rate and the growth ability were examined, and cellular morphology was assayed with DAPI fluorescent staining and Scanning electron microscope（SEM）. The possibilities of applying ADM as cells carrier or deliverer in the field of transplantation were evaluated. The result revealed that pure fibroblasts were isolated through the specific method. Skin fibroblasts could adhere to ADM easily, and the adhesion rate was 96.78%, displaying no significant difference （P〉0. 05） when compared with that rate of the control holes. The cells on the scaffolds and those on the control holes showed similar growth tendencies, but the activity of the former was lower （P〈0.01）. The integral nucleus with blue fluorescence could be observed on the ADM under fluorescence microscope. The number of fibroblasts scaled up with the cultured time, The results of SEM showed that the state of cell was good and the fibroblasts were fused into a layer after being cultured for 5-10d. So rabbit fibroblasts can attach, survive, grow and proliferate on the ADM in a healthy way. It is entirely possible to use ADM as an appropriate scaffold material for the culture of fibroblasts and as a material for transplantations.