Abstract
An improved internal gelation approach is developed to encapsulate single mammalian cells in monodisperse alginate microbeads as small as 26 μm in diameter and at rates of up to 1 kHz with high cell viability. The cell damage resulting from contact with calcium carbonate nanoparticles as gelation reagents is eliminated by employing a co-flow microfluidic device, and the cell exposure to low pH is minimized by a chemically balanced off-chip gelation step. These modifications significantly improve the viability of cells encapsulated in gelled alginate particles. Two different mammalian cell types are encapsulated with viability of over 84 %. The cells are functional and continue to grow inside the microparticles.
References
Capretto L, Mazzitelli S, Balestra C, Tosi A, Nastruzzi C (2008) Effect of the gelation process on the production of alginate microbeads by microfluidic chip technology. Lab Chip 8:617–621
Chachques JC, Trainini JC, Lago N, Masoli OH, Barisani JL, Cortes-Morichetti M, Schussler O, Carpentier A (2007) Myocardial assistance by grafting a new bioartificial upgraded myocardium (MAGNUM clinical trial): one year follow-up. Cell Transpl 16:927–934
Choi C-H, Jung J-H, Rhee Y, Kim D-P, Shim S-E, Lee C-S (2007) Generation of monodisperse alginate microbeads and in situ encapsulation of cell in microfluidic device. Biomed Microdevices 9:855–862
Drury JL, Mooney DJ (2003) Hydrogels for tissue engineering: scaffold design variables and applications. Biomaterials 24:4337–4351
Edd JF, Di Carlo D, Humphry KJ, Koster S, Irimia D, Weitz DA, Toner M (2008) Controlled encapsulation of single-cells into monodisperse picolitre drops. Lab Chip 8:1262–1264
Guo MT, Rotem A, Heyman JA, Weitz DA (2012) Droplet microfluidics for high-throughput biological assays. Lab Chip 12:2146–2155
Hoesli CA, Raghuram K, Kiang RLJ, Mocinecová D, Hu X, Johnson JD, Lacík I, Kieffer TJ, Piret JM (2011) Pancreatic cell immobilization in alginate beads produced by emulsion and internal gelation. Biotechnol Bioeng 108:424–434
Holtze C, Rowat AC, Agresti JJ, Hutchison JB, Angile FE, Schmitz CHJ, Koster S, Duan H, Humphry KJ, Scanga RA, Johnson JS, Pisignano D, Weitz DA (2008) Biocompatible surfactants for water-in-fluorocarbon emulsions. Lab Chip 8:1632–1639
Lee KY, Mooney DJ (2012) Alginate: properties and biomedical applications. Prog Polym Sci 37:106–126
Maguire T, Novik E, Schloss R, Yarmush M (2006) Alginate‐PLL microencapsulation: effect on the differentiation of embryonic stem cells into hepatocytes. Biotechnol Bioeng 93:581–591
Martinez CJ, Kim JW, Ye C, Ortiz I, Rowat AC, Marquez M, Weitz D (2012) A microfluidic approach to encapsulate living cells in uniform alginate hydrogel microparticles. Macromol Biosci 12:946–951
Mazzitelli S, Borgatti M, Breveglieri G, Gambari R, Nastruzzi C (2011) Encapsulation of eukaryotic cells in alginate microparticles: cell signaling by TNF-alpha through capsular structure of cystic fibrosis cells. J Cell Commun Signal 5:157–165
Nicodemus GD, Bryant SJ (2008) Cell encapsulation in biodegradable hydrogels for tissue engineering applications. Tissue Eng Part B Rev 14:149–165
Pierigè F, Serafini S, Rossi L, Magnani M (2008) Cell-based drug delivery. Adv Drug Deliv Rev 60:286–295
Prakash S, Chang TMS (1996) Microencapsulated genetically engineered live E. coli DH5 cells administered orally to maintain normal plasma urea level in uremic rats. Nat Med 2:883–887
Sakai S, Mu C, Kawabata K, Hashimoto I, Kawakami K (2006) Biocompatibility of subsieve‐size capsules versus conventional‐size microcapsules. J Biomed Mater Res A 78A:394–398
Schmidt JJ, Rowley J, Kong HJ (2008) Hydrogels used for cell‐based drug delivery. J Biomed Mater Res A 87A:1113–1122
Shintaku H, Kuwabara T, Kawano S, Suzuki T, Kanno I, Kotera H (2007) Micro cell encapsulation and its hydrogel-beads production using microfluidic device. Microsyst Technol 13:951–958
Slaughter BV, Khurshid SS, Fisher OZ, Khademhosseini A, Peppas NA (2009) Hydrogels in regenerative medicine. Adv Mater 21:3307–3329
Sugiura S, Oda T, Izumida Y, Aoyagi Y, Satake M, Ochiai A, Ohkohchi N, Nakajima M (2005) Size control of calcium alginate beads containing living cells using micro-nozzle array. Biomater 26:3327–3331
Sun Y, Ma X, Zhou D, Vacek I, Sun AM (1996) Normalization of diabetes in spontaneously diabetic cynomolgus monkeys by xenografts of microencapsulated porcine islets without immunosuppression. J Clin Invest 98:1417–1422
Tan WH, Takeuchi S (2007) Monodisperse alginate hydrogel microbeads for cell encapsulation. Adv Mater 19:2696–2701
Teh S-Y, Lin R, Hung L-H, Lee AP (2008) Droplet microfluidics. Lab Chip 8:198–220
Um E, Lee D-S, Pyo H-B, Park J-K (2008) Continuous generation of hydrogel beads and encapsulation of biological materials using a microfluidic droplet-merging channel. Microfluid Nanofluid 5:541–549
Workman VL, Dunnett SB, Kille P, Palmer DD (2008) On‐chip alginate microencapsulation of functional cells. Macromol Rapid Commun 29:165–170
Xu Q, Hashimoto M, Dang TT, Hoare T, Kohane DS, Whitesides GM, Langer R, Anderson DG (2009) Preparation of monodisperse biodegradable polymer microparticles using a microfluidic flow‐focusing device for controlled drug delivery. Small 5:1575–1581
Yu J, Du KT, Fang Q, Gu Y, Mihardja SS, Sievers RE, Wu JC, Lee RJ (2010) The use of human mesenchymal stem cells encapsulated in RGD modified alginate microspheres in the repair of myocardial infarction in the rat. Biomaterials 31:7012–7020
Acknowledgments
The authors thank Prof. D. A. Weitz, J. Heyman, A. Khavari, S. Utech, and R. Sperling for helpful discussions. S. Akbari, acknowledges the support of Prof. H. R. Shea and Swiss national foundation (Grant No. 200020-140394).
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Samin Akbari and Tohid Pirbodaghi have contributed equally to this article.
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Akbari, S., Pirbodaghi, T. Microfluidic encapsulation of cells in alginate particles via an improved internal gelation approach. Microfluid Nanofluid 16, 773–777 (2014). https://doi.org/10.1007/s10404-013-1264-z
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DOI: https://doi.org/10.1007/s10404-013-1264-z