In vitro co-culture of decellularized matrix of porcine colon with colon cancer HCT116 cells

doi:10.3969/j.issn.2095-4344.3049

Abstract

Abstract: BACKGROUND: Acellular scaffolds have been widely used in the study of various tumors in recent years. The spatial arrangement, biomechanical properties and biocompatibility of the scaffolds are helpful to restore the microenvironment of tumor cell growth.
OBJECTIVE: To investigate the characteristics and advantages of porcine colon acellular scaffold as an in vitro model of colon cancer.
METHODS: Acellular scaffolds of porcine colon were prepared by soaking fresh porcine colon with 2% SDS, 1% TritonX-100 and 0.5% EDTA combined with repeated shaking. Human colon cancer HCT116 cells were inoculated on the mucous surface of porcine colon acellular scaffold. The growth of cells on the scaffold was observed by live-death staining. The morphology of cells on the scaffold was observed by phalloidin staining, and the longitudinal growth of cells on the scaffold was observed by hematoxylin-eosin staining and immunofluorescence staining.
RESULTS AND CONCLUSION: (1) Live-death staining showed that on the first day of culture, HCT116 cells could well gather in the pores of the mucosal layer of the scaffolds, and few dead cells were found. On the third day, the cells gradually spread out of the pores, and a small number of cells grew and connected into pieces, and there were no dead cells. On the 7th day, the cell growth density further increased and grew into a sheet on the surface of the mucous layer, and there were no dead cells and exfoliated cells. (2) On the 7th day of culture, hematoxylin-eosin staining and immunofluorescence staining showed that some HCT116 cells could grow into clusters in the pores, and some cells continued to grow to the submucosa along the pores, showing the characteristics of invasive growth of intestinal cancer in situ. (3) On the 7th day of culture, the phalloidin staining showed that HCT116 cells were in close contact with the scaffold mucosa at the ultrastructural level, and had well-differentiated epithelial morphological characteristics. (4) The result shows that porcine colon acellular scaffold can be used as a carrier for three-dimensional culture of colon cancer cells.

Key words: materials, tumor, scaffold, acellular scaffold, three-dimensional culture, colon cancer, invasive growth

CLC Number:

Zhu Mingqi, Zhou Jingxu, Lin Lizhu, Chen Zeren, Liao Zhixiao . In vitro co-culture of decellularized matrix of porcine colon with colon cancer HCT116 cells[J]. Chinese Journal of Tissue Engineering Research, 2021, 25(10): 1533-1538.

Figures/Tables 5

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