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用于MCF-7乳腺癌细胞系体外生长的壳聚糖基质的表征与评价

Characterization and evaluation of chitosan matrix for in vitro growth of MCF-7 breast cancer cell lines.

作者信息

Dhiman H K Harpreet K, Ray A R Alok R, Panda A K Amulya K

机构信息

Centre for Biomedical Engineering, Indian Institute of Technology, Aruna Asaf Ali Marg, New Delhi-11 0016, India.

出版信息

Biomaterials. 2004 Sep;25(21):5147-54. doi: 10.1016/j.biomaterials.2003.12.025.

Abstract

Biodegradable polymer scaffolds were prepared from chitosan with varying degree of deacetylation for in vitro culture of human breast cancer MCF-7 cell lines. These polymers were characterized in terms of functional groups by FTIR and swelling properties. Polymers having high degree of deacetylation showed better swelling properties irrespective of the molecular weight. These polymers were biocompatible and non-toxic towards human epithelial MCF-7 cell lines. Attachment kinetics of MCF-7 cell lines on to polymer scaffold was investigated and it was observed that polymer having high degree of deacetylation favored better cell attachment. In CPIII polymer scaffold having 80% degree of deacetylation, a maximum of 1 millions cells per mg pf polymer were adsorbed within 1h. It appears that high swelling and high degree of deacetylation of chitosan helped in better adsorption of cancer cell lines. The cellular morphology of the attached cells on chitosan matrix was similar to that observed with regular plastic culture with the difference that, cells grew as three-dimensional clumps on chitosan matrix. Polymer having high degree of deacetylation not only favored better adsorption but also showed improved cell growth kinetics. Maximum cell concentration of 6.5 x 10(5) cells/ml was achieved in 5 days culture on CPIII polymer scaffold. The glucose consumption and lactate production pattern of the MCF-7 cell lines on chitosan polymer matrix were similar to that observed on cell growth on tissue culture flask. These results indicate that chitosan scaffold having high degree of deacetylation can be used for three-dimensional growth of MCF-7 cancer cell lines. Such in vitro 3D culture of cancer cells can thus be used as a model for the cytotoxic evaluation of anticancer drugs.

摘要

采用不同脱乙酰度的壳聚糖制备了可生物降解的聚合物支架,用于人乳腺癌MCF-7细胞系的体外培养。通过傅里叶变换红外光谱(FTIR)对这些聚合物的官能团进行了表征,并研究了其溶胀性能。无论分子量如何,脱乙酰度高的聚合物都表现出更好的溶胀性能。这些聚合物具有生物相容性,对人上皮MCF-7细胞系无毒。研究了MCF-7细胞系在聚合物支架上的附着动力学,发现脱乙酰度高的聚合物有利于更好的细胞附着。在脱乙酰度为80%的CPIII聚合物支架中,每毫克聚合物在1小时内最多可吸附100万个细胞。似乎壳聚糖的高溶胀性和高脱乙酰度有助于更好地吸附癌细胞系。壳聚糖基质上附着细胞的细胞形态与常规塑料培养中观察到的相似,不同之处在于,细胞在壳聚糖基质上以三维团块形式生长。脱乙酰度高的聚合物不仅有利于更好的吸附,还显示出改善的细胞生长动力学。在CPIII聚合物支架上培养5天后,细胞最大浓度达到6.5×10⁵个细胞/毫升。MCF-7细胞系在壳聚糖聚合物基质上的葡萄糖消耗和乳酸产生模式与在组织培养瓶中细胞生长时观察到的相似。这些结果表明,脱乙酰度高的壳聚糖支架可用于MCF-7癌细胞系的三维生长。因此,这种癌细胞的体外三维培养可作为抗癌药物细胞毒性评估的模型。

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