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Three-dimensional (3D) printing of mouse primary hepatocytes to generate 3D hepatic structure
- Source :
- Annals of Surgical Treatment and Research
- Publication Year :
- 2017
- Publisher :
- The Korean Surgical Society, 2017.
-
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 × 10 7 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.
- Subjects :
- 0301 basic medicine
Pathology
medicine.medical_specialty
Maintenance
Culture
02 engineering and technology
Alginate scaffold
Biology
Immunofluorescence
law.invention
03 medical and health sciences
law
Gene expression
medicine
Functional ability
Polymerase chain reaction
Three-dimensional printing
Primary (chemistry)
medicine.diagnostic_test
Albumin
021001 nanoscience & nanotechnology
Molecular biology
030104 developmental biology
Collagenase
Hepatocytes
Surgery
Original Article
0210 nano-technology
medicine.drug
Subjects
Details
- Language :
- English
- ISSN :
- 22886796 and 22886575
- Volume :
- 92
- Issue :
- 2
- Database :
- OpenAIRE
- Journal :
- Annals of Surgical Treatment and Research
- Accession number :
- edsair.doi.dedup.....50fb102fac946171cc9327b275cc552d