A parametric study was performed to understand the effect of preparation parameters on size, morphology, and encapsulation efficiency (i.e., cells/microcapsule) of alginate microcapsules prepared using the electrostatic spray method. The preparation parameters studied include sodium alginate concentration, spray voltage, flow rate, and cell density. It was found that both the flow rate and spray voltage have a significant impact on microcapsule size while the microcapsule morphology is greatly influenced by both the sodium alginate concentration and spray voltage. To obtain small cell-loaded microcapsules with good morphology (i.e., round in shape and uniform in size) and high encapsulation efficiency ( cells/microcapsule), the optimal ranges of spray voltage, flow rate, alginate concentration, and cell density are from 1.6–1.8 kV, 1.5–3 ml/h, >1.5% (w/v), and (3–5)106 cells/ml, respectively. Under optimal preparation conditions, cells were found to survive the microencapsulation process well.
Issue Section:
Technical Briefs
1.
Chang
, T. M. S.
, 1964, “Semipermeable Microcapsules
,” Science
0036-8075, 146
(3643
), pp. 524
–525
.2.
Lim
, F.
, and Sun
, A. M.
, 1980, “Microencapsulated Islets as Bioartificial Endocrine Pancreas
,” Science
0036-8075, 210
(4472
), pp. 908
–910
.3.
Orive
, G.
, Hernandez
, R. M.
, Rodriguez Gascon
, A.
, Calafiore
, R.
, Chang
, T. M.
, de Vos
, P.
, Hortelano
, G.
, Hunkeler
, D.
, Lacik
, I.
, and Pedraz
, J. L.
, 2004, “History, Challenges and Perspectives of Cell Microencapsulation
,” Trends Biotechnol.
0167-7799, 22
(2
), pp. 87
–92
.4.
Ding
, H. F.
, Liu
, R.
, Li
, B. G.
, Lou
, J. R.
, Dai
, K. R.
, and Tang
, T. T.
, 2007, “Biologic Effect and Immunoisolating Behavior of BMP-2 Gene-Transfected Bone Marrow-Derived Mesenchymal Stem Cells in APA Microcapsules
,” Biochem. Biophys. Res. Commun.
0006-291X, 362
(4
), pp. 923
–927
.5.
Tobias
, C. A.
, Han
, S. S.
, Shumsky
, J. S.
, Kim
, D.
, Tumolo
, M.
, Dhoot
, N. O.
, Wheatley
, M. A.
, Fischer
, I.
, Tessler
, A.
, and Murray
, M.
, 2005, “Alginate Encapsulated BDNF-Producing Fibroblast Grafts Permit Recovery of Function After Spinal Cord Injury in the Absence of Immune Suppression
,” J. Neurotrauma
0897-7151, 22
(1
), pp. 138
–156
.6.
Zhang
, H.
, Zhu
, S. J.
, Wang
, W.
, Wei
, Y. J.
, and Hu
, S. S.
, 2008, “Transplantation of Microencapsulated Genetically Modified Xenogeneic Cells Augments Angiogenesis and Improves Heart Function
,” Gene Ther.
0969-7128, 15
(1
), pp. 40
–48
.7.
Visted
, T.
, Bjerkvig
, R.
, and Enger
, P. O.
, 2001, “Cell Encapsulation Technology as a Therapeutic Strategy for CNS Malignancies
,” J. Neuro-Oncol.
0167-594X, 3
(3
), pp. 201
–210
.8.
Tu
, J.
, Bolla
, S.
, Barr
, J.
, Miedema
, J.
, Li
, X.
, and Jasti
, B.
, 2005, “Alginate Microparticles Prepared by Spray-Coagulation Method: Preparation, Drug Loading and Release Characterization
,” Int. J. Pharm.
0378-5173, 303
(1–2
), pp. 171
–181
.9.
Orive
, G.
, Tam
, S. K.
, Pedraz
, J. L.
, and Halle
, J. P.
, 2006, “Biocompatibility of Alginate-Poly-L-Lysine Microcapsules for Cell Therapy
,” Biomaterials
0142-9612, 27
(20
), pp. 3691
–3700
.10.
Rinaudo
, M.
, 2008, “Main Properties and Current Applications of Some Polysaccharides as Biomaterials
,” Polym. Int.
0959-8103, 57
(3
), pp. 397
–430
.11.
Halle
, J. P.
, Leblond
, F. A.
, Pariseau
, J. F.
, Jutras
, P.
, Brabant
, M. J.
, and Lepage
, Y.
, 1994, “Studies on Small (<300 Microns) Microcapsules: II–Parameters Governing the Production of Alginate Beads by High Voltage Electrostatic Pulses
,” Cell Transplant.
, 3
(5
), pp. 365
–372
. 0963-689712.
Amsden
, B. G.
, and Goosen
, M. F. A.
, 1997, “An Examination of Factors Affecting the Size, Distribution and Release Characteristics of Polymer Microbeads Made Using Electrostatics
,” J. Controlled Release
0168-3659, 43
(2–3
), pp. 183
–196
.13.
Uludag
, H.
, De Vos
, P.
, and Tresco
, P. A.
, 2000, “Technology of Mammalian Cell Encapsulation
,” Adv. Drug Delivery Rev.
0169-409X, 42
(1–2
), pp. 29
–64
.14.
Hartman
, R. P. A.
, Brunner
, D. J.
, Camelot
, D. M. A.
, Marijnissen
, J. C. M.
, and Scarlett
, B.
, 2000, “Jet Break-Up in Electrohydrodynamic Atomization in the Cone-Jet Mode
,” J. Aerosol Sci.
0021-8502, 31
(1
), pp. 65
–95
.15.
Strand
, B. L.
, Gaserod
, O.
, Kulseng
, B.
, Espevik
, T.
, and Skjak-Braek
, G.
, 2002, “Alginate-Polylysine-Alginate Microcapsules: Effect of Size Reduction on Capsule Properties
,” J. Microencapsul.
0265-2048, 19
(5
), pp. 615
–630
.16.
Li
, B. G.
, Hua
, T. C.
, Zhang
, H. D.
, Wang
, Y. F.
, and Wang
, G. X.
, 2002, “Cryopreservation and Xenotransplantation Studies of Microencapsulated Rat Pancreatic Islets
,” Cryo Letters
, 23
(1
), pp. 47
–54
. 0143-204417.
Koch
, S.
, Schwinger
, C.
, Kressler
, J.
, Heinzen
, C.
, and Rainov
, N. G.
, 2003, “Alginate Encapsulation of Genetically Engineered Mammalian Cells: Comparison of Production Devices, Methods and Microcapsule Characteristics
,” J. Microencapsul.
0265-2048, 20
(3
), pp. 303
–316
.18.
Bugarski
, B.
, Li
, Q. L.
, Goosen
, M. F. A.
, Poncelet
, D.
, Neufeld
, R. J.
, and Vunjakg
, G.
, 1994, “Electrostatic Droplet Generation—Mechanism of Polymer Droplet Formation
,” AIChE J.
0001-1541, 40
(6
), pp. 1026
–1031
.19.
Zhou
, Y.
, Sun
, T.
, Chan
, M.
, Zhang
, J.
, Han
, Z.
, Wang
, X.
, Toh
, Y.
, Chen
, J. P.
, and Yu
, H.
, 2005, “Scalable Encapsulation of Hepatocytes by Electrostatic Spraying
,” J. Biotechnol.
0168-1656, 117
(1
), pp. 99
–109
.20.
Xie
, J.
, and Wang
, C. H.
, 2007, “Electrospray in the Dripping Mode for Cell Microencapsulation
,” J. Colloid Interface Sci.
0021-9797, 312
(2
), pp. 247
–255
.21.
Goosen
, M. F. A.
, AlGhafri
, A. S.
, ElMardi
, O.
, AlBelushi
, M. I. J.
, AlHajri
, H. A.
, Mahmoud
, E. S. E.
, and Consolacion
, E. C.
, 1997, “Electrostatic Droplet Generation for Encapsulation of Somatic Tissue: Assessment of High-Voltage Power Supply
,” Biotechnol. Prog.
8756-7938, 13
(4
), pp. 497
–502
.22.
Klokk
, T. I.
, and Melvik
, J. E.
, 2002, “Controlling the Size of Alginate Gel Beads by Use of a High Electrostatic Potential
,” J. Microencapsul.
0265-2048, 19
(4
), pp. 415
–424
.23.
Sugiura
, S.
, Oda
, T.
, Izumida
, Y.
, Aoyagi
, Y.
, Satake
, M.
, Ochiai
, A.
, Ohkohchi
, N.
, and Nakajima
, M.
, 2005, “Size Control of Calcium Alginate Beads Containing Living Cells Using Micro-Nozzle Array
,” Biomaterials
0142-9612, 26
(16
), pp. 3327
–3331
.24.
Shintaku
, H.
, Kuwabara
, T.
, Kawano
, S.
, Suzuki
, T.
, Kanno
, I.
, and Kotera
, H.
, 2007, “Micro Cell Encapsulation and Its Hydrogel-Beads Production Using Microfluidic Device
,” Microsystem Technologies-Micro-and Nanosystems-Information Storage and Processing Systems
, 13
(8–10
), pp. 951
–958
.25.
Huang
, K. S.
, Liu
, M. K.
, Wu
, C. H.
, Yen
, Y. T.
, and Lin
, Y. C.
, 2007, “Calcium Alginate Microcapsule Generation on a Microfluidic System Fabricated Using the Optical Disk Process
,” J. Micromech. Microeng.
0960-1317, 17
(8
), pp. 1428
–1434
.26.
Ross
, C. J.
, and Chang
, P. L.
, 2002, “Development of Small Alginate Microcapsules for Recombinant Gene Product Delivery to the Rodent Brain
,” J. Biomater. Sci., Polym. Ed.
0920-5063, 13
(8
), pp. 953
–962
.27.
Sugiura
, S.
, Oda
, T.
, Aoyagi
, Y.
, Matsuo
, R.
, Enomoto
, T.
, Matsumoto
, K.
, Nakamura
, T.
, Satake
, M.
, Ochiai
, A.
, Ohkohchi
, N.
, and Nakajima
, M.
, 2007, “Microfabricated Airflow Nozzle for Microencapsulation of Living Cells Into 150 Micrometer Microcapsules
,” Biomed. Microdevices
1387-2176, 9
(1
), pp. 91
–99
.28.
Wilson
, J. T.
, and Chaikof
, E. L.
, 2008, “Challenges and Emerging Technologies in the Immunoisolation of Cells and Tissues
,” Adv. Drug Delivery Rev.
0169-409X, 60
(2
), pp. 124
–145
.29.
Sakai
, S.
, Kawabata
, K.
, Ono
, T.
, Ijima
, H.
, and Kawakami
, K.
, 2004, “Preparation of Mammalian Cell-Enclosing Subsieve-Sized Capsules (<100 Micron) in a Coflowing Stream
,” Biotechnol. Bioeng.
0006-3592, 86
(2
), pp. 168
–173
.30.
Chin Heng
, B.
, Yu
, H.
, and Chye Ng
, S.
, 2004, “Strategies for the Cryopreservation of Microencapsulated Cells
,” Biotechnol. Bioeng.
0006-3592, 85
(2
), pp. 202
–213
.31.
He
, X.
, Park
, E. Y.
, Fowler
, A.
, Yarmush
, M. L.
, and Toner
, M.
, 2008, “Vitrification by Ultra-Fast Cooling at a Low Concentration of Cryoprotectants in a Quartz Micro-Capillary: A Study Using Murine Embryonic Stem Cells
,” Cryobiology
0011-2240, 56
(3
), pp. 223
–232
.32.
Toner
, M.
, Cravalho
, E. G.
, and Karel
, M.
, 1990, “Thermodynamics and Kinetics of Intracellular Ice Formation During Freezing of Biological Cells
,” J. Appl. Phys.
0021-8979, 67
(3
), pp. 1582
–1593
.33.
Karlsson
, J. O. M.
, Cravalho
, E. G.
, and Toner
, M.
, 1994, “A Model of Diffusion-Limited Ice Growth Inside Biological Cells During Freezing
,” J. Appl. Phys.
0021-8979, 75
(9
), pp. 4442
–4445
.34.
Berejnov
, V.
, Husseini
, N. S.
, Alsaied
, O. A.
, and Thorne
, R. E.
, 2006, “Effects of Cryoprotectant Concentration and Cooling Rate on Vitrification of Aqueous Solutions
,” J. Appl. Crystallogr.
0021-8898, 39
, pp. 244
–251
.35.
Zhang
, Y.
, Wang
, W.
, Xie
, Y.
, Yu
, W.
, Lv
, G.
, Guo
, X.
, Xiong
, Y.
, and Ma
, X.
, 2008, “Optimization of Microencapsulated Recombinant CHO Cell Growth, Endostatin Production, and Stability of Microcapsule in Vivo
,” J. Biomed. Mater. Res., Part B: Appl. Biomater.
1552-4973, 84b
(1
), pp. 79
–88
.36.
Zhang
, W. J.
, Li
, B. G.
, Zhang
, C.
, Xie
, X. H.
, and Tang
, T. T.
, 2008, “Biocompatibility and Membrane Strength of C3H10T1/2 Cell-Loaded Alginate-Based Microcapsules
,” Cytotherapy
, 10
(1
), pp. 90
–97
. 1465-324937.
Oh
, I.
, Ozaki
, M.
, Miyazato
, A.
, Sato
, K.
, Meguro
, A.
, Muroi
, K.
, Nagai
, T.
, Mano
, H.
, and Ozawa
, K.
, 2007, “Screening of Genes Responsible for Differentiation of Mouse Mesenchymal Stromal Cells by DNA Micro-Array Analysis of C3H10T1/2 and C3H10T1/2-Derived Cell Lines
,” Cytotherapy
, 9
(1
), pp. 80
–90
. 1465-324938.
Cai
, A. H.
, Chen
, H.
, An
, W. K.
, Li
, X. S.
, and Zhou
, Y.
, 2008, “Optimization of Composition and Technology for Phosphate Graphite Mold
,” Mater. Des.
0264-1275, 29
(9
), pp. 1835
–1839
.39.
Snedecor
, G. W.
, and Cochran
, W. G.
, 1989, Statistical Methods
, Iowa State University
, Ames, IA
.40.
NIST
, “NIST/SEMATECH E-Handbook of Statistical Methods
,” http://www.itl.nist.gov/div898/handbook/http://www.itl.nist.gov/div898/handbook/.41.
Arus
, L.
, Orive
, G.
, Hernandez
, R.
, Rodriguez
, A.
, Rojas
, A.
, and Pedraz
, J. L.
, 2005, “The Influence of Cellular Seeding Density in the Microencapsulation of Hybridoma Cells
,” J. Biomater. Sci., Polym. Ed.
0920-5063, 16
(4
), pp. 521
–529
.42.
Qi
, W. T.
, Zhang
, Y.
, Ma
, J. A.
, Guo
, X.
, Xie
, Y. B.
, Wang
, W.
, and Ma
, X. J.
, 2006, “Optimization of the Cell Seeding Density and Modeling of Cell Growth and Metabolism Using the Modified Gompertz Model for Microencapsulated Animal Cell Culture
,” Biotechnol. Bioeng.
0006-3592, 93
(5
), pp. 887
–895
.Copyright © 2009
by American Society of Mechanical Engineers
You do not currently have access to this content.
