AIMS Bioengineering, 2017, 4(4): 402-417. doi: 10.3934/bioeng.2017.4.402.

Research article

Export file:

Format

  • RIS(for EndNote,Reference Manager,ProCite)
  • BibTex
  • Text

Content

  • Citation Only
  • Citation and Abstract

Development of an automated chip culture system with integrated on-line monitoring for maturation culture of retinal pigment epithelial cells

1 Department of Biotechnology, Graduate School of Engineering, Osaka University, 2-1 Yamadaoka, Suita, Osaka 565-0871, Japan
2 Shimadzu Co., 1, Nishinokyo Kuwabara-cho, Nakagyo-ku, Kyoto 604-8511, Japan
3 Micronix, Inc., 24-1, Shinarami, Tai, Kuse-gun, Kumiyama-cho 613-0036, Japan

In cell manufacturing, the establishment of a fully automated, microfluidic, cell culture system that can be used for long-term cell cultures, as well as for process optimization is highly desirable. This study reports the development of a novel chip bioreactor system that can be used for automated long-term maturation cultures of retinal pigment epithelial (RPE) cells. The system consists of an incubation unit, a medium supply unit, a culture observation unit, and a control unit. In the incubation unit, the chip contains a closed culture vessel (2.5 mm diameter, working volume 9.1 μL), which can be set to 37 °C and 5% CO2, and uses a gas-permeable resin (poly- dimethylsiloxane) as the vessel wall. RPE cells were seeded at 5.0 × 104 cells/cm2 and the medium was changed every day by introducing fresh medium using the medium supply unit. Culture solutions were stored either in the refrigerator or the freezer, and fresh medium was prepared before any medium change by warming to 37 °C and mixing. Automated culture was allowed to continue for 30 days to allow maturation of the RPE cells. This chip culture system allows for the long-term, bubble-free, culture of RPE cells, while also being able to observe cells in order to elucidate their cell morphology or show the presence of tight junctions. This culture system, along with an integrated on-line monitoring system, can therefore be applied to long-term cultures of RPE cells, and should contribute to process control in RPE cell manufacturing.
  Figure/Table
  Supplementary
  Article Metrics

Keywords retinal pigment epithelium; chip cell culture system; tight junction formation; maturation culture; on-line monitoring

Citation: Mee-Hae Kim, Masakazu Inamori, Masakazu Akechi, Hirohisa Abe, Yoshiki Yagi, Masahiro Kino-oka. Development of an automated chip culture system with integrated on-line monitoring for maturation culture of retinal pigment epithelial cells. AIMS Bioengineering, 2017, 4(4): 402-417. doi: 10.3934/bioeng.2017.4.402

References

  • 1. Da CL, Chen FK, Ahmado A, et al. (2007) RPE transplantation and its role in retinal disease. Prog Retin Eye Res 26: 598–635.    
  • 2. Tezel TH, Priore LVD, Berger AS, et al. (2007) Adult retinal pigment epithelial transplantation in exudative age-related macular degeneration. Am J Ophthalmol 143: 584–395.    
  • 3. Uygun BE, Sharma N, Yarmush M (2009) Retinal pigment epithelium differentiation of stem cells: current status and challenges. Crit Rev Biomed Eng 37: 355–375.    
  • 4. Dang Y, Zhang C, Zhu Y (2015) Stem cell therapies for age-related macular degeneration: the past, present, and future. Clin Interv Aging 10: 255–264.
  • 5. Melville H, Carpiniello M, Hollis K, et al. (2013) Stem cells: a new paradigm for disease modeling and developing therapies for age-related macular degeneration. J Transl Med 11: 53.    
  • 6. Fields MA, Bowrey HE, Gong J, et al. (2015) Retinoid processing in induced pluripotent stem cell-derived retinal pigment epithelium cultures. Prog Mol Biol Transl Sci 134: 477–490.    
  • 7. Strauss O (2005) The retinal pigment epithelium in visual function. Physiol Rev 85: 845–881.    
  • 8. Fronk AH, Vargis E (2016) Methods for culturing retinal pigment epithelial cells: a review of current protocols and future recommendations. J Tissue Eng 7: 1–23.
  • 9. Burke JM, Skumatz CM, Irving PE, et al. (1996) Phenotypic heterogeneity of retinal pigment epithelial cells in vitro and in situ. Exp Eye Res 62: 63–73.    
  • 10. McKay BS, Irving PE, Skumatz CM, et al. (1997) Cell-cell adhesion molecules and the development of an epithelial phenotype in cultured human retinal pigment epithelial cells. Exp Eye Res 65: 661–671.    
  • 11. Sonoi R, Kim MH, Kino-oka M (2016) Locational heterogeneity of maturation by changes in migratory behaviors of human retinal pigment epithelial cells in culture. J Biosci Bioeng 119: 107–112.
  • 12. Sonoi R, Kim MH, Kino-oka M (2016) Facilitation of uniform maturation of human retinal pigment epithelial cells through collective movement in culture. J Biosci Bioeng 121: 220–226.    
  • 13. Rothbauer M, Wartmann D, Charwat V, et al. (2015) Recent advances and future applications of microfluidic live-cell microarrays. Biotechnol Adv 33: 948–961.    
  • 14. Yi C, Li CW, Ji S, et al. (2006) Microfluidics technology for manipulation and analysis of biological cells. Anal Chim Acta 560: 1–23.    
  • 15. Tam J, Cordier GA, Bálint Š, et al. (2014) A microfluidic platform for correlative live-cell and super-resolution microscopy. PLoS One 9: e115512.    
  • 16. Kartalov EP, Anderson WF, Scherer A (2006) The analytical approach to polydimethylsiloxane microfluidic technology and its biological applications. J Nanosci Nanotechno 6: 2265–2277.    
  • 17. Sung JH, Shuler ML (2009) Prevention of air bubble formation in a microfluidic perfusion cell culture system using a microscale bubble trap. Biomed Microdevices 11: 731–738.    
  • 18. Zhang W, Lin S, Wang C, et al. (2009) PMMA/PDMS valves and pumps for disposable microfluidics. Lab Chip 9: 3088–3094.    
  • 19. Leclerc E, Sakai Y, Fujii T (2004) Microfluidic PDMS (polydimethylsiloxane) bioreactor for large-scale culture of hepatocytes. Biotechnol Progr 20: 750–755.    
  • 20. Ren KN, Zhou JH, Wu HK (2013) Materials for microfluidic chip fabrication. Accounts Chem Res 46: 2396–2406.    
  • 21. Gómez-Sjöberg R, Leyrat AA, Pirone DM, et al. (2007) Versatile, fully automated, microfluidic cell culture system. Anal Chem 79: 8557–8563.    
  • 22. Wartmann D, Rothbauer M, Kuten O, et al. (2015) Automated, miniaturized and integrated quality control-on-chip (qc-on-a-chip) for advanced cell therapy applications. Front Mater 2: 60.
  • 23. Jaccard N, Macown RJ, Super A, et al. (2014) Automated and online characterization of adherent cell culture growth in a microfabricated bioreactor. J Lab Autom 19: 437–443.    
  • 24. Bahnemann J, Rajabi N, Fuge G, et al. (2013) A new integrated lab-on-a-chip system for fast dynamic study of mammalian cells under physiological conditions in bioreactor. Cells 2: 349–360.    
  • 25. Kawaguchi R, Yu J, Honda J, et al. (2007) A membrane receptor for retinol binding protein mediates cellular uptake of vitamin A. Science 315: 820–825.    
  • 26. Bok D (1990) Processing and transport of retinoids by the retinal pigment epithelium. Eye 4: 326–332.    
  • 27. Bise R, Kanade T, Yin Z, et al. (2011) Automatic cell tracking applied to analysis of cell migration in wound healing assay. International Conference of the IEEE Engineering in Medicine and Biology Society, 6174–6179.
  • 28. Dorfer M, Kazmar T, Šmíd M, et al. (2016) Associating approximate paths and temporal sequences of noisy detections: application to the recovery of spatio-temporal cancer cell trajectories. Med Image Anal 27: 72–83.    
  • 29. Ker DF, Weiss LE, Junkers SN, et al. (2011) An engineered approach to stem cell culture: automating the decision process for real-time adaptive subculture of stem cells. PLoS One 6: e27672.    

 

Reader Comments

your name: *   your email: *  

Copyright Info: 2017, Masahiro Kino-oka, et al., licensee AIMS Press. This is an open access article distributed under the terms of the Creative Commons Attribution Licese (http://creativecommons.org/licenses/by/4.0)

Download full text in PDF

Export Citation

Copyright © AIMS Press All Rights Reserved