Spherical and cylindrical microencapsulation of living cells using microfluidic devices

  • Hong, Joung-Sook (Department of Biomedical Engineering, Korea University) ;
  • Shin, Su-Jung (Department of Biomedical Engineering, Korea University) ;
  • Lee, Sang-Hoon (Department of Biomedical Engineering, Korea University) ;
  • Wong, Edeline (Australian Institute for Bioengineering and Nanotechnology, The University of Queensland) ;
  • Cooper-White, Justin (Australian Institute for Bioengineering and Nanotechnology, The University of Queensland)
  • Published : 2007.11.30

Abstract

Microencapsulation of cells within microfluidic devices enables explicit control of the membrane thickness or cell density, resulting in improved viability of the transplanted cells within an aggressive immune system. In this study, living cells (3T3 and L929 fibroblast cells) are encapsulated within a semi-permeable membrane (calcium crosslinked alginate gel) in two different device designs, a flow focusing and a core-annular flow focusing geometry. These two device designs produce a bead and a long microfibre, respectively. For the alginate bead, an alginate aqueous solution incorporating cells flows through a flow focusing channel and an alginate droplet is formed from the balance of interfacial forces and viscous drag forces resulting from the continuous (oil) phase flowing past the alginate solution. It immediately reacts with an adjacent $CaCl_2$ drop that is extruded into the main flow channel by another flow focusing channel downstream of the site of alginate drop creation. Depending on the flow conditions, monodisperse microbeads of sizes ranging from $50-200\;{\mu}m$ can be produced. In the case of the microfibre, the alginate solution with cells is extruded into a continuous phase of $CaCl_2$ solution. The diameter of alginate fibres produced via this technique can be tightly controlled by changing both flow rates. Cell viability in both forms of alginate encapsulant was confirmed by a LIVE/DEAD cell assay for periods of up to 24 hours post encapsulation.

Keywords

References

  1. Anna, S.L., N. Bontoux and H.A. Stone, 2003, Formation of dispersions using 'flow focusing' in microchannels, App. Phy. Lett. 82(3), 364-366 https://doi.org/10.1063/1.1537519
  2. Chang, T.M.S., 1964, Semipermeable microcapsule, Science 146, 524-525 https://doi.org/10.1126/science.146.3643.524
  3. Fan, M.Y., Z.P. Lum, X.W. Fu, L. Levesque, I.T. Tai and A.M. Sun, 1990, Reversal of diabetes in BB rats by transplantation of encapsulated pancreatic islets, Diabetes 39, 519-522 https://doi.org/10.2337/diabetes.39.4.519
  4. Hong, J.S. and Cooper-White, J.J., 2007, Drop formation of a non-Newtonian fluid in a flow-focusing microfluidic channel, in preparation
  5. Jeong, W.J., J.Y. Kim, S.J. Kim, S.H. Lee, G. Mensing, and D.J. Beebe, 2004, Lab Chip 4, 576-580 https://doi.org/10.1039/b411249k
  6. Jeong, W.J., J.Y. Kim, J. Choo, E.K. Lee, C.S. Han, D.J. Beebe, G.H. Seong and S.H. Lee, 2005, Langmuir 21, 3738-3741 https://doi.org/10.1021/la050105l
  7. Khattak, S.F., S.R. Bhatia and S.C. Roberts, 2005, Pluronic F127 as a cell encapsulation material: utilization of membrane-stabilizing agents, Tissue Engineering 11(5/6), 974-983 https://doi.org/10.1089/ten.2005.11.974
  8. Lacy, P.E. and M. Kostianovsky, 1967, Method for the isolation of intact islets of Langerhans from the rat pancreas. Diabetes 16, 35-39 https://doi.org/10.2337/diab.16.1.35
  9. Li, C.H., 2006, Microfluidic Lab-on a-chip for chemical and biological analysis and discovery, CRC press
  10. Lim, F. and A.M. Sun, 1980, Microencapsulated islets as bioartificial endocrine pancreas, Science 210, 908-910 https://doi.org/10.1126/science.6776628
  11. Microprobes, Product information MP07013, LIVE/DEAD Reduced Biohazard Viability/Cytotoxicity Kit#1 (L-7013)
  12. Microprobes, Product information MP03224, LIVE/DEAD Viability/Cytotoxicity Kit for mammalian cells
  13. Rayleigh, L., 1879, On the capillary phenomena of jets, Proc. R. Soc. 29, 71-97
  14. Strand, B.L., 2001, Poly-L-lysine induces fibrosis on alginate microcapsules via the induction of cytokines, Cell Transplant. 10, 263-275 https://doi.org/10.3727/000000001783986800
  15. Sun, Y.L., X.J. Ma, D.B. Zhou, I. Vacek and A.M. Sun, 1996, Normalization of diabetes in spontaneously diabetic cynomologous monkeys by xenografts of microencapsulated porcine islets without immunosuppression, J. Clin. Invest. 98, 1417-1422 https://doi.org/10.1172/JCI118929
  16. Sugiura, S., T. Oda, Y. Izumida, Y. Aoyagi, M. Satake, A. Ochiai, N. Ohkohchi and M. Nakajima, 2005, Size control of calcium alginate beads containing living cells using micro-nozzle array, Biomaterials 26, 3327-3331 https://doi.org/10.1016/j.biomaterials.2004.08.029
  17. Utada, A.S., E. Lorenceau, D.R. Link, P.D. Kaplan, H.A. Stone and D.A. Weitz, 2005, Monodisperse double emulsions generated from a microcapillary device, Science 308, 537-541 https://doi.org/10.1126/science.1109164
  18. Xu, Q., M. Nakajima, 2004, The generation of highly monodisperse droplets through the breakup of hydrodynamically focused microthread in a microfluidic device, App. Phy. Lett. 85(17), 3726-3728 https://doi.org/10.1063/1.1812380