J Korean Soc Transplant.  2010 Jun;24(2):101-109. 10.4285/jkstn.2010.24.2.101.

Alginate Microencapsulation of Islet Cells Using Electrostatic Droplet Generator

Affiliations
  • 1Department of Biological Science, Gachon University of Medicine and Science, Incheon, Korea. jykim85@gachon.ac.kr
  • 2Transplantation Research Institute, Department of Internal Medicine, Seoul National University College of Medicine, Seoul, Korea.

Abstract

BACKGROUND
Microencapsulation of islet cells has been studied for the treatment of type I diabetes to protect islets from immune attack by recipient cells during islet transplantation. In this study, we established optimal preparation conditions for islet microcapsules with good morphology and cell viability by employing an electrostatic droplet generator.
METHODS
To obtain good quality islet microcapsules, various parameters such as the inner diameter of the electrostatic droplet generator nozzle, concentrations and infusion rates of alginate, electrostatic strength, and calcium chloride concentrations were tested. The size and shape of the capsules and cell viability were examined by light microscopy, and insulin secretion from the cells was determined by an ELISA analysis.
RESULTS
The optimal preparation conditions for microencapsulation were a 0.35 mm inner nozzle diameter, a 1.75% alginate concentration, a 20 mL/hr alginate infusion rate, a 5 kV electrostatic potential, and a 75 mM calcium chloride. Under these conditions, over 90% of the capsules had a proper size (300~350 micrometer) and shape, and cell viability was 91%. Cell viability was maintained at greater than 80% even after the capsules were cultured for 2 weeks. However, glucose-induced insulin secretion of encapsulated islet cells was reduced by 85% compared to that of nonencapsulated cells.
CONCLUSIONS
The results showed that an encapsulation technique using an electrostatic droplet generator is useful for making islet microcapsules with good morphology and cell viability. This technique is necessary to study microencapsulation using primary islets, enhancing glucose-induced insulin secretion, and to perform a functional evaluation of encapsulated islets in vivo in the near future.

Keyword

Microencapsulation; Islet cells; Alginate; Electrostatic potential

MeSH Terms

Alginates
Calcium Chloride
Capsules
Cell Survival
Drug Compounding
Enzyme-Linked Immunosorbent Assay
Glucuronic Acid
Hexuronic Acids
Insulin
Islets of Langerhans
Islets of Langerhans Transplantation
Light
Microscopy
Alginates
Calcium Chloride
Capsules
Glucuronic Acid
Hexuronic Acids
Insulin

Reference

References

1). Kabelitz D, Geissler EK, Soria B, Schroeder IS, Fandrich F, Chatenoud L. Toward cell-based therapy of type I diabetes. Trends Immunol. 2008; 29:68–74.
Article
2). O'Sullivan ES, Johnson AS, Omer A, Hollister-Lock J, Bonner-Weir S, Colton CK, et al. Rat islet cell aggregates are superior to islets for transplantation in microcapsules. Diabetologia. 2010; 53:9370–45.
3). Bonner-Weir S, Weir GC. New sources of pancreatic be-ta-cells. Nat Biotechnol. 2005; 23:857–61.
4). Mallett AG, Korbutt GS. Alginate modification improves longterm survival and function of transplanted encapsulated islets. Tissue Eng Part A. 2009; 15:1301–9.
Article
5). Lim F, Sun AM. Microencapsulated islets as bioartificial endocrine pancreas. Science. 1980; 210:908–10.
Article
6). Kizilel S, Garfinkel M, Opara E. The bioartificial pancreas: progress and challenges. Diabetes Technol Ther. 2005; 7:968–85.
Article
7). Cornolti R, Figliuzzi M, Remuzzi A. Effect of micro- and macroencapsulation on oxygen consumption by pancreatic islets. Cell Transplant. 2009; 18:195–201.
Article
8). de Vos P, Hamel AF, Tatarkiewicz K. Considerations for successful transplantation of encapsulated pancreatic islets. Diabetologia. 2002; 45:159–73.
Article
9). Lim F. Microencapsulation of living mammalian cells. Adv Biotechnol Processes. 1988; 7:185–97.
10). Duvivier-Kali VF, Omer A, Parent RJ, O'Neil JJ, Weir GC. Complete protection of islets against allorejection and autoimmunity by a simple barium alginate membrane. Diabetes. 2001; 50:1698–705.
11). Dusseault J, Tam SK, Menard M, Polizu S, Jourdan G, Yahia L, et al. Evaluation of alginate purification methods: effect on polyphenol, endotoxin, and protein contamination. J Biomed Mater Res A. 2006; 76:243–51.
Article
12). Orive G, Hernandez RM, Rodriguez Gascon A, Calafiore R, Chang TM, de Vos P, et al. History, challenges and perspectives of cell microencapsulation. Trends Biotechnol. 2004; 22:87–92.
Article
13). de Vos P, Faas MM, Strand B, Calafiore R. Alginate-based microcapsules for immunoisolation of pancreatic islets. Biomaterials. 2006; 27:5603–17.
Article
14). Kurachi M, Nakashima T, Miyajima C, Iwamoto Y, Muramatsu T, Yamaguchi K, et al. Comparison of the activities of various alginates to induce TNF-alpha secretion in RAW264.7 cells. J Infect Chemothe. 2005; 11:199–203.
15). Santerre RF, Cook RA, Crisel RM, Sharp JD, Schmidt RJ, Williams DC, et al. Insulin synthesis in a clonal cell line of simian virus 40-transformed hamster pancreatic beta cells. Proc Natl Acad Sci USA. 1981; 78:4339–43.
Article
16). Okajima F, Kurihara M, Ono C, Nakajima Y, Tanimura K, Sugihara H, et al. Oxidized but not acetylated low-density lipoprotein reduces preproinsulin mRNA expression and secretion of insulin from HIT-T15 cells. Biochim Biophys Acta. 2005; 1687:173–80.
Article
17). Wolters GH, Fritschy WM, Gerrits D, van Schilfgaarde R. A versatile alginate droplet generator applicable for microencapsulation of pancreatic islets. J Appl Biomater. 1991; 3:281–6.
Article
18). Lim DJ, Cho JH, Choi YH, Park HS, Hong OK, Kwon HS, et al. Optimization of microencapsulation of the islets. Tissue Eng Regen Med. 2005; 2:280–6. (임동준, 조재형, 최윤희, 박헌석, 홍옥기, 권혁상, 등. 췌도 미세캡슐의 최적화. 한국조직공학재생의학회지 2005;2: 280–6.).
19). Koo SK, Kim SC, Wee YM, Kim JH, Kim YH, Jung EJ, et al. Experimental micro encapsulation of pancreatic islets with air-driven droplet generator and alginate. J Korean Soc Transplant. 2007; 21:38–48. (구선경, 김송철, 위유미, 김진희, 김양희, 정은정, 등. 공기분사형 캡슐제조기와 알지네이트를 이용한 췌장 도세포의 미세캡슐화에 대한 초기 실험. 대한이식학회지 2007;21: 38–48.).
20). Klokk TI, Melvik JE. Controlling the size of alginate gel beads by use of a high electrostatic potential. J Microencapsul. 2002; 19:415–24.
Article
21). Lewiń ska D, Rosiń ski S, Weryń ski A. Influence of process conditions during impulsed electrostatic droplet formation on size distribution of hydrogel beads. Artif Cells Blood Substit Immobil Biotechnol. 2004; 32:41–53.
22). Rong L, Baoguo L, Huifeng D, Tingting T. Factors influencing the formation of MSCs APA microcapsules. Conf Proc IEEE Eng Med Biol Soc. 2005; 5:4900–3.
23). Zhang WJ, Li BG, Zhang C, Xie XH, Tang TT. Biocompatibility and membrane strength of C3H10T1/2 cell-loaded alginate-based microcapsules. Cytotherapy. 2008; 10:90–7.
Article
24). Dufrane D, Goebbels RM, Saliez A, Guiot Y, Gianello P. Six-month survival of microencapsulated pig islets and alginate biocompatibility in primates: proof of concept. Transplantation. 2006; 81:1345–53.
Article
25). Darrabie MD, Kendall WF, Opara EC. Effect of alginate composition and gelling cation on microbead swelling. J Microencapsul. 2006; 23:613–21.
Article
26). Baruch L, Benny O, Gilert A, Ukobnik M, Ben Itzhak O, Machluf M. Alginate-PLL cell encapsulation system Co-entrapping PLGA-microspheres for the continuous release of anti-inflammatory drugs. Biomed Microdevices. 2009; 11:1103–13.
Article
27). Sandler S, Andersson A, Eizirik DL, Hellerstrom C, Espevik T, Kulseng B, et al. Assessment of insulin secretion in vitro from microencapsulated fetal porcine is-let-like cell clusters and rat, mouse, and human pancreatic islets. Transplantation. 1997; 63:1712–8.
28). Omer A, Duvivier-Kali V, Fernandes J, Tchipashvili V, Colton CK, Weir GC. Long-term normoglycemia in rats receiving transplants with encapsulated islets. Transplantation. 2005; 79:52–8.
Article
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