Three-dimensional (3D) printing of mouse primary hepatocytes to generate 3D hepatic structure.
10.4174/astr.2017.92.2.67
- Author:
Yohan KIM
1
;
Kyojin KANG
;
Jaemin JEONG
;
Seung Sam PAIK
;
Ji Sook KIM
;
Su A PARK
;
Wan Doo KIM
;
Jisun PARK
;
Dongho CHOI
Author Information
1. Department of Surgery, Hanyang University College of Medicine, Seoul, Korea. crane87@hanyang.ac.kr
- Publication Type:Original Article
- Keywords:
Hepatocytes;
Three-dimensional printing;
Culture;
Maintenance
- MeSH:
Animals;
Collagenases;
Drug Evaluation, Preclinical;
Fluorescent Antibody Technique;
Gene Expression;
Hepatocytes*;
Liver;
Liver, Artificial;
Methods;
Mice*;
Printing, Three-Dimensional;
Real-Time Polymerase Chain Reaction
- From:Annals of Surgical Treatment and Research
2017;92(2):67-72
- CountryRepublic of Korea
- Language:English
-
Abstract:
PURPOSE: The major problem in producing artificial livers is that primary hepatocytes cannot be cultured for many days. Recently, 3-dimensional (3D) printing technology draws attention and this technology regarded as a useful tool for current cell biology. By using the 3D bio-printing, these problems can be resolved. METHODS: To generate 3D bio-printed structures (25 mm × 25 mm), cells-alginate constructs were fabricated by 3D bio-printing system. Mouse primary hepatocytes were isolated from the livers of 6–8 weeks old mice by a 2-step collagenase method. Samples of 4 × 10⁷ hepatocytes with 80%–90% viability were printed with 3% alginate solution, and cultured with well-defined culture medium for primary hepatocytes. To confirm functional ability of hepatocytes cultured on 3D alginate scaffold, we conducted quantitative real-time polymerase chain reaction and immunofluorescence with hepatic marker genes. RESULTS: Isolated primary hepatocytes were printed with alginate. The 3D printed hepatocytes remained alive for 14 days. Gene expression levels of Albumin, HNF-4α and Foxa3 were gradually increased in the 3D structures. Immunofluorescence analysis showed that the primary hepatocytes produced hepatic-specific proteins over the same period of time. CONCLUSION: Our research indicates that 3D bio-printing technique can be used for long-term culture of primary hepatocytes. It can therefore be used for drug screening and as a potential method of producing artificial livers.
