Skip to main content
SpringerLink
Log in

Development of microfluidic alginate microbead generator tunable by pulsed airflow injection for the microencapsulation of cells

  • Research Paper
  • Published:
Microfluidics and Nanofluidics Aims and scope Submit manuscript

Abstract

Recently, microbead generation and microencapsulation of cells using microfluidic technology have been actively pursued for various applications. However, most of the proposed systems are not only technically demanding, but might also be harmful to the encapsulated cells. To tackle these issues, this study reports a microfluidic alginate microbead generator consisting of a polydimethylsiloxane (PDMS) microfluidic chip and an integrated quartz microcapillary tube. The working principle is based on the use of a pulsed airflow to segment a continuous alginate suspension flow to form suspension fragments in a microchannel and then alginate microbeads when they were delivered out the microfluidic system to a sterile calcium chloride solution through a microcapillary tube. In this study, the alginate suspension fragments with varied sizes in the microchannel can be generated either by modulating the alginate suspension flow rate or the pulsation frequency of airflow injection. By fine tuning the size of them, the alginate microbeads can be generated in a size-controllable manner. Results showed that alginate microbeads with the size ranging from 150 to 370 μm in diameter can be generated at the suspension flow rate and airflow injection frequency ranges of 2–4 μl/min and 0.6–35 Hz, respectively. Besides, the alginate microbeads generated by the system were tested with excellent size uniformity (CV: 3.1–5.1%). Moreover, its application for the microencapsulation of chondrocytes in alginate microbeads was also demonstrated with high cell viability (96 ± 2%). As a whole, the proposed device has paved an alternative route to perform alginate microbead generation or microencapsulation of cells in a simple, continuous, controllable, uniform, cell friendly, and less contaminated manner.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5
Fig. 6
Fig. 7
Fig. 8

Similar content being viewed by others

References

  • Abbah SA, Lu WW, Chan D, Cheung KMC, Liu WG, Zhao F, Li ZY, Leong JCY, Luk KDK (2006) In vitro evaluation of alginate encapsulated adipose-tissue stromal cells for use as injectable bone graft substitute. Biochem Biophys Res Commun 347:185–191

    Article  Google Scholar 

  • Al-Rubeai M, Singh RP, Goldman MH, Emery AN (1995) Death mechanisms of animal cells in conditions of intensive agitation. Biotechnol Bioeng 45:463–472

    Article  Google Scholar 

  • Cai X, Lin Y, Ou G, Luo E, Man Y, Yuan Q, Gong P (2007) Ectopic osteogenesis and chondrogenesis of bone marrow stromal stem cells in alginate system. Cell Biol Int 31:776–783

    Article  Google Scholar 

  • Choi CH, Jung JH, Rhee YW, Kim DP, Shim SE, Lee CS (2007) Generation of monodisperse alginate microspheres and in situ encapsulation of cells in microfluidic device. Biomed Microdevices 9:855–862

    Article  Google Scholar 

  • Christopher GF, Anna SL (2007) Microfluidic methods for generating continuous droplet streams. J Phys D Appl Phys 40:R319–R336

    Article  Google Scholar 

  • Chung C, Mesa J, Miller GJ, Randolph MA, Gill TJ, Burdick JA (2006) Effects of articular chondrocyte expansion on neocartilage formation in photocrosslinked hyaluronic acid networks. Tissue Eng 12:2665–2673

    Article  Google Scholar 

  • Clausell-Tormos J, Lieber D, Baret JC, El-Harrak A, Miller OJ, Frenz L, Blouwolff J, Humphry KJ, Koster S, Duan H, Holtze C, Weitz DA, Griffths AD, Merten CA (2008) Droplet-based microfluidic platforms for the encapsulation and screening of mammalian cells and multicellular organisms. Chem Biol 15:427–437

    Article  Google Scholar 

  • Dvir-Ginzberg M, Konson A, Cohen S, Agbaria R (2007) Entrapment of retroviral vector producer cells in three-dimensional alginate scaffolds for potential use in cancer gene therapy. J Biomed Mater Res B Appl Biomater 80:59–66

    Google Scholar 

  • Elliott RB, Escobar L, Tan PLJ, Garkavenko O, Calafiore R, Basta P, Vasconcellos AV, Emerich DF, Thanos C, Bambra C (2005) Intraperitoneal alginate-encapsulated neonatal porcine islets in a placebo-controlled study with 16 diabetic cynomolgus primates. Transplant Proc 37:3505–3508

    Article  Google Scholar 

  • Emerich DF, Thanos CG, Goddard M, Skinner SJM, Geany MS, Bell WJ, Bintz B, Schneider P, Chu Y, Babu RS, Borlongan CV, Boekelheide K, Hall S, Bryant B, Kordower JH (2006) Extensive neuroprotection by choroid plexus transplants in excitotoxin lesioned monkeys. Neurobiol Dis 23:471–480

    Article  Google Scholar 

  • Gaver D, Kute S (1998) Theoretical model study of the influence of fluid stresses on a cell adhering to a microchannel wall. Biophys J 75:721–733

    Article  Google Scholar 

  • Gunther A, Khan SA, Thalmann M, Trachsel F, Jensen KF (2004) Transport and reaction in microscale segmented gas–liquid flow. Lab Chip 4:278–286

    Article  Google Scholar 

  • Huang KS, Liu MK, Wu CH, Yen YT, Lin YC (2007) Calcium alginate microcapsule generation on a microfluidic system fabricated using the optical disk process. J Micromech Microeng 17:1428–1434

    Article  Google Scholar 

  • Huang SB, Wu MH, Cui ZF, Cui Z, Lee GB (2008) A membrane-based serpentine-shape pneumatic micropump with pumping performance modulated by fluidic resistance. J Micromech Microeng 18:045008

    Article  Google Scholar 

  • Jork A, Thurmer F, Cramer H, Zimmermann G, Gessner P, Hamel K, Hofmann G, Kuttler B, Hahn HJ, Josimovic-Alasevic O, Fritsch KG, Zimmermann U (2002) Biocompatible alginate from freshly collected laminaria pallida for implantation. Appl Microbiol Biotechnol 53:224–229

    Article  Google Scholar 

  • Kaigler D, Krebsbach PH, Wang Z, West ER, Horger K, Mooney DJ (2006) Transplanted endothelial cells enhance orthotopic bone regeneration. J Dent Res 85:633–637

    Article  Google Scholar 

  • Klock G, Pfeffermann A, Ryser C, Grohn P, Kuttler B, Hahn HJ, Zimmermann U (1997) Biocompatibility of mannuronic acid-rich alginates. Biomaterials 18:707–713

    Article  Google Scholar 

  • Koster S, Angile FE, Duan H, Agresti JJ, Wintner A, Schmitz C, Rowat AC, Merten CA, Pisignnano D, Griffiths AD, Weitz DA (2008) Drop-based microfluidic devices for encapsulation of single cells. Lab Chip 8:1110–1115

    Article  Google Scholar 

  • Lin YH, Chen CT, Hung LLH, Lee GB (2007) Multiple-channel emulsion chips utilizing pneumatic choppers for biotechnology applications. Biomed Microdevices 9:833–843

    Article  Google Scholar 

  • Liu VA, Bhatia SN (2002) Three-dimensional photopatterning of hydrogels containing living cells. Biomed Microdevices 4:257–266

    Article  Google Scholar 

  • Liu K, Ding HJ, Liu J, Chen Y, Zhao XZ (2006) Shape-controlled production of biodegradable calcium alginate gel microparticles using a novel microfluidic device. Langmuir 22:9453–9457

    Article  Google Scholar 

  • Morimoto Y, Tan WH, Takeuchi S (2009) Three-dimensional axisymmetric flow-focusing device using stereolithography. Biomed Microdevices 11:369–377

    Article  Google Scholar 

  • Murua A, Portero A, Orive G, Hernandez RM, Castro MD, Pedraz JL (2008) Cell microencapsulation technology: towards clinical application. J Control Release 132:76–83

    Article  Google Scholar 

  • Schneider S, Feilen PJ, Brunnenmeier F, Minnemann T, Zimmermann H, Zimmermann U, Weber MM (2005) Long-term graft function of adult rat and human islets encapsulated in novel alginate-based microcapsules after transplantation in immunocompetent diabetic mice. Diabetes 54:687–693

    Article  Google Scholar 

  • 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

    Article  Google Scholar 

  • Song H, Bringer MR, Tice JD, Gerdts CJ, Ismagilov RF (2003) Experimental test of scaling of mixing by chaotic advection in droplets moving through microfluidic channels. Appl Phys Lett 83:4664–4666

    Article  Google Scholar 

  • Srinivasan V, Pamula VK, Fair RB (2004) An integrated digital microfluidic lab-on-a-chip for clinical diagnostics on human physiological fluids. Lab Chip 4:310–315

    Article  Google Scholar 

  • Stathopoulos NA, Hellums JD (2004) Shear stress effects on human embryonic kidney cells in vitro. Biotechnol Bioeng 27:1021–1026

    Article  Google Scholar 

  • Sugihara-Seki M (2000) Flow around cells adhered to a microvessel wall I. Fluid stresses and forces acting on the cells. Biorheology 37:341–359

    Google Scholar 

  • Sugihara-Seki M (2001) Flow around cells adhered to a microvessel wall II. Comparison to flow around adherent cells in channel flow. Biorheology 38:3–13

    Google Scholar 

  • Sugiura S, Oda T, Aoyagi Y, Matsuo R, Enomoto T, Matsumoto K, Nakamura T, Satake M, Ochiai A, Ohkohchi N, Nakajima M (2007) Microfabricated airflow nozzle for microencapsulation of living cells into 150 micrometers microcapsules. Biomed Microdevices 9:91–99

    Article  Google Scholar 

  • Tan WH, Takeuchi S (2007) Monodisperse alginate hydrogel microbeads for cell encapsulation. Adv Mater 19:2696–2701

    Article  Google Scholar 

  • Teh SY, Lin R, Hung LS, Lee AP (2008) Droplet microfluidics. Lab Chip 8:198–220

    Article  Google Scholar 

  • Wu MH, Urban JPG, Cui Z, Cui ZF (2006) Development of PDMS microbioreactor with well-defined and homogenous culture environment for chondrocyte 3-D culture. Biomed Microdevices 8:331–340

    Article  Google Scholar 

  • Wu MH, Urban JPG, Cui ZF, Cui Z, Xu X (2007) Effect of extracellular pH on matrix synthesis by chondrocytes in 3D agarose gel. Biotechnol Prog 23:430–434

    Article  Google Scholar 

  • Wu MH, Huang SB, Cui ZF, Cui Z, Lee GB (2008) Development of perfusion-based micro 3-D cell culture platform and its application for high throughput drug testing. Sens Actuators B 129:231–240

    Article  Google Scholar 

  • Xie J, Wang CH (2007) Electrospray in the dipping mode for cell microencapsulation. J Colloid Interface Sci 312:247–255

    Article  Google Scholar 

  • Xu X, Urban JPG, Tirlapur U, Wu MH, Cui Z, Cui ZF (2006) Influence of perfusion on metabolism and matrix production by bovine articular chondrocytes in hydrogel scaffolds. Biotechnol Bioeng 93:1103–1111

    Article  Google Scholar 

  • Xu J, Li S, Tan J, Luo G (2008) Controllable preparation of monodispersed calcium alginate microbeads in a novel microfluidic system. Chem Eng Technol 31:1223–1226

    Article  Google Scholar 

  • Yang CH, Huang KS, Chang JY (2007) Manufacturing monodisperse chitosan microparticles containing ampicillin using a microchannel chip. Biomed Microdevices 9:253–259

    Article  Google Scholar 

  • Yeh J, Lin Y, Karp JM, Gantz J, Chandawarkar A, Eng G (2006) Micromolding of shape-controlled, harvestable cell-laden hydrogels. Biomaterials 27:5391–5398

    Article  Google Scholar 

Download references

Acknowledgment

This project is sponsored by the National Science Council (NSC) in Taiwan (NSC 98-2221-E-182-008).

Author information

Authors and Affiliations

  1. Graduate Institute of Biochemical and Biomedical Engineering, Chang Gung University, Taoyuan, Taiwan, ROC

    Min-Hsien Wu & Wei-Chung Pan

Authors
  1. Min-Hsien Wu
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Wei-Chung Pan
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Min-Hsien Wu.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Wu, MH., Pan, WC. Development of microfluidic alginate microbead generator tunable by pulsed airflow injection for the microencapsulation of cells. Microfluid Nanofluid 8, 823–835 (2010). https://doi.org/10.1007/s10404-009-0522-6

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s10404-009-0522-6

Keywords

Search

Navigation