Development of high throughput microfluidic cell culture chip for perfusion 3-dimensional cell culture-based chemosensitivity assay

Abstract This study reports a microfluidic cell culture chip encompassing 36 microbioreactors for high throughput perfusion 3-dimensional (3D) cell culture-based chemosensitivity assays. Its advantages include the capability for multiplexed medium delivery, and the function for both efficient and high throughput micro-scale 3D culture construct preparation and loading. The results showed that the proposed medium pumping mechanism was able to provide a uniform pumping rates ranging from 1.2 to 3.9 μl h −1 . In addition, the simple cell/hydrogel loading scheme has been proven to be able to carry out 3D cell culture construct preparation and loading precisely and efficiently. Furthermore, a chemosensitivity assay was successfully demonstrated using the proposed cell culture chip. The results obtained were also compared with the same evaluation based on a conventional 2D monolayer cell culture. It can be concluded that the choice of cell culture format can result in different chemosensitivity evaluation results. Overall, because of the nature of miniaturized perfusion 3D cell culture, the cell culture chip not only can provide stable, well-defined and more biologically relevant culture environments, but it also features low consumption of research resources. All these traits are found particularly useful for high-precision and high-throughput 3D cell culture-based assays.

[1]  A. Chiu,et al.  Enhanced transformation and chemosensitivity of NIH3T3 cells transduced with hepatoma up-regulated protein. , 2006, Biochemical and biophysical research communications.

[2]  Christopher S. Chen,et al.  Cells lying on a bed of microneedles: An approach to isolate mechanical force , 2003, Proceedings of the National Academy of Sciences of the United States of America.

[3]  K. Jensen,et al.  Cells on chips , 2006, Nature.

[4]  Mina J Bissell,et al.  Modeling dynamic reciprocity: engineering three-dimensional culture models of breast architecture, function, and neoplastic transformation. , 2005, Seminars in cancer biology.

[5]  Ying Zhang,et al.  Development of an in Vitro Multicellular Tumor Spheroid Model Using Microencapsulation and Its Application in Anticancer Drug Screening and Testing , 2008, Biotechnology progress.

[6]  Samuel K Sia,et al.  Real-time microfluidic system for studying mammalian cells in 3D microenvironments. , 2008, Analytical chemistry.

[7]  Min-Hsien Wu.  Simple poly(dimethylsiloxane) surface modification to control cell adhesion , 2009 .

[8]  Matsuhiko Nishizawa,et al.  Multi-channel 3-D cell culture device integrated on a silicon chip for anticancer drug sensitivity test. , 2005, Biomaterials.

[9]  W W Minuth,et al.  Artificial Tissues in Perfusion Culture , 1997, The International journal of artificial organs.

[10]  Jr-Lung Lin,et al.  Application of indium tin oxide (ITO)-based microheater chip with uniform thermal distribution for perfusion cell culture outside a cell incubator , 2010, Biomedical microdevices.

[11]  Kenneth M. Yamada,et al.  Taking Cell-Matrix Adhesions to the Third Dimension , 2001, Science.

[12]  B. Lin,et al.  Cell-based high content screening using an integrated microfluidic device. , 2007, Lab on a chip.

[13]  Joo H. Kang,et al.  Microfluidic biomechanical device for compressive cell stimulation and lysis , 2007 .

[14]  A. Craft,et al.  Cytogenetic abnormalities of small round cell tumours. , 1994, Medical and pediatric oncology.

[15]  D. Dobrota,et al.  In vitro assays for the evaluation of drug resistance in tumor cells , 2009, Clinical and Experimental Medicine.

[16]  Shuichi Shoji,et al.  Development of a poly-dimethylsiloxane microfluidic device for single cell isolation and incubation , 2009 .

[17]  Chun-jian Qi,et al.  In vitro chemosensitivity in breast cancer using ATP-tumor chemosensitivity assay , 2009, Archives of pharmacal research.

[18]  Michal Chudy,et al.  PDMS/glass microfluidic cell culture system for cytotoxicity tests and cells passage , 2010 .

[19]  F. Pampaloni,et al.  The third dimension bridges the gap between cell culture and live tissue , 2007, Nature Reviews Molecular Cell Biology.

[20]  Ralf Ehret,et al.  Real-time assessment of cytotoxicity by impedance measurement on a 96-well plate , 2007 .

[21]  N. Aronson,et al.  Chemotherapy sensitivity and resistance assays: a systematic review. , 2004, Journal of clinical oncology : official journal of the American Society of Clinical Oncology.

[22]  Zheng Cui,et al.  Simple surface treatments to modify protein adsorption and cell attachment properties within a poly(dimethylsiloxane) micro‐bioreactor , 2006 .

[23]  Alok R Ray,et al.  Three-dimensional chitosan scaffold-based MCF-7 cell culture for the determination of the cytotoxicity of tamoxifen. , 2005, Biomaterials.

[24]  Feng Xu,et al.  Three-dimensional polymeric systems for cancer cell studies , 2007, Cytotechnology (Dordrecht).

[25]  F. Guilak,et al.  Monolayer cell expansion conditions affect the chondrogenic potential of adipose‐derived stem cells , 2008, Biotechnology and bioengineering.

[26]  Minseok S. Kim,et al.  Microvalve‐assisted patterning platform for measuring cellular dynamics based on 3D cell culture , 2008, Biotechnology and bioengineering.

[27]  A. Abbott.  Cell culture: Biology's new dimension , 2003, Nature.

[28]  A. Štanclová,et al.  Human epithelial growth factor receptor 2[Ile655Val] polymorphism and risk of breast fibroadenoma , 2008, European journal of cancer prevention : the official journal of the European Cancer Prevention Organisation.

[29]  D. Beebe,et al.  Microenvironment design considerations for cellular scale studies. , 2004, Lab on a chip.

[30]  O. Orwar,et al.  Microfluidic gradient-generating device for pharmacological profiling. , 2005, Analytical chemistry.

[31]  J. Urban,et al.  Development of PDMS microbioreactor with well-defined and homogenous culture environment for chondrocyte 3-D culture , 2006, Biomedical microdevices.

[32]  Gwo-Bin Lee,et al.  Microfluidic cell culture systems for drug research. , 2010, Lab on a chip.

[33]  Zheng Cui,et al.  Effect of Extracellular pH on Matrix Synthesis by Chondrocytes in 3D Agarose Gel , 2007, Biotechnology progress.

[34]  Gwo-Bin Lee,et al.  A high throughput perfusion-based microbioreactor platform integrated with pneumatic micropumps for three-dimensional cell culture , 2008, Biomedical microdevices.

[35]  P. Benya,et al.  Dedifferentiated chondrocytes reexpress the differentiated collagen phenotype when cultured in agarose gels , 1982, Cell.

[36]  Taisuke Masuda,et al.  Three-dimensional cell culture device utilizing thin membrane deformation by decompression , 2010 .

[37]  Gwo-Bin Lee,et al.  Development of perfusion-based micro 3-D cell culture platform and its application for high throughput drug testing , 2008 .

[38]  Zheng Cui,et al.  A membrane-based serpentine-shape pneumatic micropump with pumping performance modulated by fluidic resistance , 2008 .

[39]  Laurent Griscom,et al.  Trends in the development of microfluidic cell biochips for in vitro hepatotoxicity. , 2007, Toxicology in vitro : an international journal published in association with BIBRA.

[40]  S. Quake,et al.  Monolithic microfabricated valves and pumps by multilayer soft lithography. , 2000, Science.

[41]  Hossein Baharvand,et al.  Differentiation of human embryonic stem cells into hepatocytes in 2D and 3D culture systems in vitro. , 2006, The International journal of developmental biology.

[42]  Gwo-Bin Lee,et al.  Automatic bio-sampling chips integrated with micro-pumps and micro-valves for disease detection. , 2005, Biosensors & bioelectronics.

[43]  C. Kwak,et al.  Enhanced chemosensitivity of bladder cancer cells to cisplatin by suppression of clusterin in vitro. , 2004, Cancer letters.

[44]  S. Sahoo,et al.  3-D tumor model for in vitro evaluation of anticancer drugs. , 2008, Molecular pharmaceutics.