Authors have no conflicts of interest to disclose.
Suspension culture of hepatocyte-derived reporter cells in presence of albumin to form stable three-dimensional spheroids
Article first published online: 2 MAY 2013
Copyright © 2013 Wiley Periodicals, Inc.
Biotechnology and Bioengineering
Volume 110, Issue 9, pages 2548–2555, September 2013
How to Cite
Weeks, C. A., Newman, K., Turner, P. A., Rodysill, B., Hickey, R. D., Nyberg, S. L. and Janorkar, A. V. (2013), Suspension culture of hepatocyte-derived reporter cells in presence of albumin to form stable three-dimensional spheroids. Biotechnol. Bioeng., 110: 2548–2555. doi: 10.1002/bit.24899
- Issue published online: 23 JUL 2013
- Article first published online: 2 MAY 2013
- Accepted manuscript online: 8 MAR 2013 01:26PM EST
- Manuscript Accepted: 28 FEB 2013
- Manuscript Revised: 25 FEB 2013
- Manuscript Received: 14 JAN 2013
- School of Dentistry Intramural Research Support Program
- National Science Foundation. Grant Number: 1033525
- 3D cell culture;
- bioartificial liver assist devices;
Several studies in the past have formed 3-dimensional (3D) spheroids of primary hepatocytes in suspension culture. Unfortunately, primary hepatocytes in a suspension environment tend to lose their differentiated function over time, generally due to damage from fluid shear stress and eventual spheroid settling. We have therefore created a novel suspension culture system, by seeding H35 rat hepatoma cells, a hepatocyte-derived cell line, in a 24-well tissue culture polystyrene (TCPS) plate placed atop an orbital shaker to create 3D spheroids. To provide stability to the formed spheroids, we used a long-chain polymer, bovine serum albumin (BSA), dissolved in the cell culture medium and/or coated on TCPS surfaces placed in suspension configurations. Our results demonstrate that BSA coating of culture surfaces resulted in uniform and well-defined spheroids with little spheroid settling or “flattening” of cell colonies in either static or suspension configurations. In BSA-coated suspension systems, spheroid size scaled with the amount of BSA dissolved in culture medium. In static uncoated cultures, the normalized rat albumin production levels were enhanced by addition of BSA within culture medium. Thus, both addition of BSA to culture medium and application of BSA as a surface coating appear to be meaningful avenues for tailoring spheroid morphology and function. This 24-well plate suspension culture system may be a valuable tool for high throughput investigations of liver cell behavior in a stable, uniform, 3D spheroid state. Biotechnol. Bioeng. 2013; 110:2548–2555. © 2013 Wiley Periodicals, Inc.