采用超临界二氧化碳技术制备多孔复合细胞支架材料；利用该支架制成细胞填充床生物反应器，在满足体内肿瘤细胞生长和维持肿瘤细胞活性所需的生理条件下，三维立体培养肿瘤细胞；在模拟体内药物与肿瘤细胞受体作用的微环境下，使药物分子与肿瘤细胞作用；固相萃取作用后的药物分子，并通过比较与活细胞和固化细胞作用后的药物色谱指纹图谱，确定与细胞特异性作用的活性药物分子，利用已知抗肿瘤和非抗肿瘤药物建立抗肿瘤药物细胞筛选模型，并应用于中草药中与肿瘤细胞特异性作用的活性成分筛选。进一步建立细胞填充床反应器-固相萃取-高效液相色谱/质谱（CPBBR-SPE-HPLC/MS）在线筛选平台及分析检测系统，实现肿瘤细胞三维立体培养、细胞特异性作用中药活性成分筛选、分离富集、成分分析与化合物结构鉴定的集成。该研究旨在发展体内细胞作用的化学物质的分离模式、实现中药复杂体系中与细胞特异性作用活性成分组的有效筛选和选择性分析。

Some porous composite scaffolds will be prepared with supercrtical CO2 saturation technology and a cell packed-bed bioreactor will be fabricated. The bioreactor can realize three-dimensional culture of cancer cells at the in vivo physiology conditions for cells to grow and maintain their viabilities. Drugs interact with cancer cells at a simulant in vivo microenvironment of cell-drug interaction, and uninteracted drugs are enriched on a column by solid phase extraction. The specific binding components will be distinguished by comparing the peak areas of the biological fingerprinting chromatograms of drugs after interacting with live and fixed cancer cells. A cell screening model for anticancer drugs will be established with well-known anticancer drugs and non-anticancer drugs, and be empolyed to screen the bioactive components interacted with cancer cells specifically from herb medicines. An online screening, analysis and detection system, cell packed-bed bioreactor coupled with solid phase extraction and HPLC/MS (CPBBR-SPE-HPLC/MS), will be further constructed. This system can accomplish three-dimensional culture of cancer cells, and screening, separation, enrichment, composition analysis and chemical structure identification of bioactive components interacted with cancer cells specifically from herb medicines. This study aimed to develop a chemical separation mode and to create an effective screening and selective analysis method for bioactive component groups interacted with cancer cells specifically from complex system of traditional Chinese medicines.