|
KMID : 0371420170920020067
|
|
Annals of Surgical Treatment and Research
2017 Volume.92 No. 2 p.67 ~ p.72
|
|
|
Three-dimensional (3D) printing of mouse primary hepatocytes to generate 3D hepatic structure
|
|
Kim Yo-Han
Kang Kyo-Jin
Jeong Jae-Min
Paik Seung-Sam
Kim Ji-Sook
Park Su-A
Kim Wan-Doo
Park Ji-Sun
Choi Dong-Ho
|
|
|
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.
|
|
|
KEYWORD
|
|
|
Hepatocytes, Three-dimensional printing, Culture, Maintenance
|
|
|
FullTexts / Linksout information
|
|
|
|
|
Listed journal information
|
|
   
|