Guan Linbo, Dan Weihua, Lin Hai, Dan Nianhua, Wang Kunyu, Liao Longli, Li Zhiqiang, Chen Min, Zeng Rui
The Key Laboratory of Leather Chemistry and Engineering of Ministry of Education, Sichuan University, Chengdu 610065, China.
Sheng Wu Yi Xue Gong Cheng Xue Za Zhi. 2009 Oct;26(5):1010-5.
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.
研究了成纤维细胞在脱细胞真皮基质(ADM)上的生长情况。从成年新西兰兔皮肤中分离出成纤维细胞,进行体外培养并随后鉴定。将细胞作为种子接种到ADM上后,检测其黏附率和生长能力,并用DAPI荧光染色和扫描电子显微镜(SEM)检测细胞形态。评估了ADM在移植领域作为细胞载体或递送剂的可能性。结果表明,通过特定方法分离出了纯成纤维细胞。皮肤成纤维细胞能够轻松黏附于ADM,黏附率为96.78%,与对照孔的黏附率相比无显著差异(P>0.05)。支架上的细胞与对照孔中的细胞显示出相似的生长趋势,但前者的活性较低(P<0.01)。在荧光显微镜下可在ADM上观察到带有蓝色荧光的完整细胞核。成纤维细胞数量随培养时间增加。SEM结果显示,培养5 - 10天后细胞状态良好,成纤维细胞融合成一层。因此,兔成纤维细胞能够以健康的方式在ADM上附着、存活、生长和增殖。将ADM用作成纤维细胞培养的合适支架材料以及移植材料是完全可行的。