CAJ | 학술논문

目的:以人肝细胞系HepG2作为种子细胞,建立一种细胞支架评价方法,为肝组织工程筛选合适支架.方法:将HepG2细胞接种在生物可降解聚合物支架--聚乳酸-乙醇酸共聚物(PLGA)、壳聚糖(3%CS)和丝素(2%SF)上,体外常规培养;采用四唑盐(MTT)比色法、HE染色和尿素氮检测试剂盒对接种在支架上HepG2细胞的生长、分布及功能情况进行检测.结果:培养在3种支架上的HepG2细胞均维持增殖状态,相比之下,丝素支架上的细胞增殖较快,而PLGA和壳聚糖支架上的细胞增殖相对缓慢;培养至第7 d时,组织学检测显示丝素支架上的HepG2细胞分布均匀,教量最多;而在PLGA和壳聚糖支架上只能发现少量细胞;细胞功能检测显示丝素和PLGA支架上的尿素合成能力下降缓慢,而壳聚糖支架上的尿素合成能力快速下降.结论:3种支架均具有比较好的生物相容性;相比较而言,丝素更适合作为肝组织工程支架;该方法可用于批量筛选肝组织工程支架.
목적:이인간세포계HepG2작위충자세포,건립일충세포지가평개방법,위간조직공정사선합괄지가.방법:장HepG2세포접충재생물가강해취합물지가--취유산-을순산공취물(PLGA)、각취당(3%CS)화사소(2%SF)상,체외상규배양;채용사서염(MTT)비색법、HE염색화뇨소담검측시제합대접충재지가상HepG2세포적생장、분포급공능정황진행검측.결과:배양재3충지가상적HepG2세포균유지증식상태,상비지하,사소지가상적세포증식교쾌,이PLGA화각취당지가상적세포증식상대완만;배양지제7 d시,조직학검측현시사소지가상적HepG2세포분포균균,교량최다;이재PLGA화각취당지가상지능발현소량세포;세포공능검측현시사소화PLGA지가상적뇨소합성능력하강완만,이각취당지가상적뇨소합성능력쾌속하강.결론:3충지가균구유비교호적생물상용성;상비교이언,사소경괄합작위간조직공정지가;해방법가용우비량사선간조직공정지가.
Obiective To set up a method of scaffold evaluation using human cell line as seed cells and screen appropriate scaffold for live tissue engineering, Methods HepG2 cells were plated onto biodegradable polymer scaffolds: PLGA, 3% chitosan (3%CS) and 2% silk fibroin (2%SF), and cultured in vitro. The growth, distribution and function of HepG2 cells in the scaffolds were evaluated using MTT assay, H.E. staining, and urea assay kit. Results HepG2 cells plated on the three scaffolds maintained a proliferative state. In contrast, the cells on the 2%SF proliferated strongly, while the cells on the PLGA and chitin proliferated poorly. Histological examination showed that HepG2 cells distributed evenly on the 2%SF scaffold with a high amount, while few cells could be found on the PLGA and ehitin at day 7. Cell function assay showed that HepG2 cells on the 2%SF and PLGA exhibited slower decrease of urea synthesis compared to HepG2 cells on the chitosan. Conclusion The three scaffolds have good biocompatibility. In contrast, 2%SF scaffold is more appropriate for liver tissue engineering. This method may be used for scale screening of scaffolds for liver tissue engineering.