Co-culture mode of tissue-engineered tracheal epithelial cells and fibroblasts: Comparison with routine culture
- VernacularTitle:构建组织工程气管上皮细胞和成纤维细胞联合培养的模式:与常规培养的比较
- Author:
Lin WANG
;
Wei JIANG
- Publication Type:Journal Article
- From:
Chinese Journal of Tissue Engineering Research
2007;0(15):-
- CountryChina
- Language:Chinese
-
Abstract:
AIM: To establish the co-culture mode of tissue-engineered tracheal seeding cells, and compare with the routine culture, so as to provide the fundament of tissue-engineered trachea construction by cell compound materials. METHODS: The experiment was completed at the laboratory, Department of Respiratory Diseases, Tangdu Hospital, the Fourth Military Medical University of Chinese PLA from May 2006 to May 2007.①Three healthy New Zealand male rabbits, weighed (250.00?0.75) g, were used in this study.②Co-culture mode: The tracheal epithelial cells and fibroblasts were isolated and then co-cultured for 7-10 days. The two kinds of cells were distinguished according to their different tolerances to trypsin. Subsequently, A hole received 0.5 g/L trypsinization, cells were suspended in DMEM medium containing 0.05 volume fraction of CO2 and then transplanted into B hole (fibroblasts); again, A hole was added with 2.5 g/L trypsin for digestion, afterwards cells were suspended with K-FSM medium and transplanted into C hole (tracheal epithelial cells). Routine culture: after isolation, tracheal epithelial cells were cultured and fibroblasts were purified, respectively.③The co-cultured cells and the traditionally cultured cells were compared via cell growth curve and cell proliferation detected by 3-(4,5-dimethylthiazo-2-yl)-2,5-diphenyl-tetrazolium bromide (MTT) assay. RESULTS: ①Cell growth: Co-cultured cells grew well, those in A hole increased intercellular apace after trypsinization, while cells in B hole were shaped as fusiform, without the manifestation of typical road stone-like cells, and cells in C hole were apposite, showing lamellar road stone-like morphology.②Cell growth curve: The growth curve of co-cultured cells was identical with that of cells by routine culture method.③Cell proliferation: MTT detection revealed no significant difference in the proliferation of co-cultured cells and cells cultured by traditional method (P
