SpringerLink
Log in

Towards the development of a bioartificial pancreas: Immunoisolation and NMR monitoring of mouse insulinomas

  • Published:
Cytotechnology Aims and scope Submit manuscript

Abstract

A promising method for diabetes treatment is the implantation of immunoisolated cells secreting insulin in response to glucose. Cell availability limits the application of this approach at a medically-relevant scale. We explore the use of transformed cells that can be grown to large homogeneous populations in develo** artificial pancreatic tissues. We also investigate the use of NMR in evaluating, non-invasively, cellular bioenergetics in the tissue environment. The system employed in this study consisted of mouse insulinoma βTC3 cells entrapped in calcium alginate/poly-L-lysine (PPL)/alginate beads. The PPL layer imposed a molecular weight cutoff of approximately 60 kDa, allowing nutrients and insulin to diffuse through but excluding high molecular weight antibodies and cytotoxic cells of the host. We fabricated a radiofrequency coil that can be double-tuned to1H and31P, and an NMR-compatible perfusion bioreactor and support circuit that can maintain cells viable during prolonged studies. The bioreactor operated differentially, was macroscopically homogeneous and allowed the acquisition of1H images and31P NMR spectra in reasonable time intervals. Results indicated that entrapment had little effect on cell viability; that insulin secretion from beads was responsive to glucose; and that the bioenergetics of perfused, entrapped cells were not grossly different from those of cells never subjected to the immobilization procedure. These findings offer promise for develo** an artificial pancreatic tissue for diabetes treatment based on continuous cell lines.

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

Access this article

Log in via an institution

Subscribe and save

Springer+ Basic
EUR 32.99 /Month
  • Get 10 units per month
  • Download Article/Chapter or Ebook
  • 1 Unit = 1 Article or 1 Chapter
  • Cancel anytime
Subscribe now

Buy Now

Price includes VAT (Germany)

Instant access to the full article PDF.

Institutional subscriptions

Similar content being viewed by others

References

  • Calafiore R (1992) Transplantation of microencapsulated pancreatic human islets for therapy of diabetes mellitus. ASAIO J. 38: 34–37.

    Google Scholar 

  • Carroll P (1992) Anatomy and physiology of islets of Langerhans. In: Ricordi C (ed.) Pancreatic Islet Cell Transplantation, pp. 7–18. Austin: R.G. Landes Co.

    Google Scholar 

  • Chang L-H, Chew WM, Weinstein PR, and James TL (1987) A balanced-matched double tuned probe for in vivo1H and31P NMR. J. Magn. Reson. 72: 168–172.

    Google Scholar 

  • Clayton HA, London NJM, Bell PRF and James RFL (1992) The transplantation of encapsulated islets of Langerhans into the peritoneal cavity of the biobreeding rat. Transplantation 54: 558–560.

    Google Scholar 

  • Constantinidis I, Chatham JC, Wehrle JP and Glickson JD (1991)In vivo 13C-NMR studies of the RIF-1 tumor after glucose administration. Magn. Res. Med. 20: 17–26.

    Google Scholar 

  • Constantinidis I, Long RC Jr, Erickson M and Sambanis A (1994) Towards the development of artificial endocrine tissues: II.31P NMR spectroscopic studies of entrapped insulin-secreting cells. Submitted.

  • Darquy S and Reach G (1985) Immunoisolation of pancreatic B cells by microencapsulation. Anin vitro study. Diabetologia 28: 776–780.

    Google Scholar 

  • Dyken JJ and Sambanis A (1994) Ammonium selectively inhibits the regulated pathway of protein secretion in two endocrine cell lines. Enz. Microb. Technol. 16: 90–98.

    Google Scholar 

  • Efrat S, Leiser M, Surana M, Tal M, Fusco-Demane D and Fleischer N (1993) Murine insulinoma cell line with normal glucoseregulated insulin secretion. Diabetes 42: 901–907.

    Google Scholar 

  • Efrat S, Linde S, Kofod H, Spector D, Delannoy M, Grant S, Hanahan D and Baekkeskov S (1988) Beta-cell lines derived from transgenic mice expressing a hybrid insulin gene-oncogene. Proc. Natl. Acad. Sci. USA 85: 9037–9041.

    Google Scholar 

  • Evanochko WT, Sakai TT, Ng TC, Krishna NR, Kim HD, Zeidler RB, Ghanta VK, Brockman RW, Schiffer LM, Braunschweiger PG and Glickson JD (1984) NMR studies ofin vivo RIF-1 tumors. Analysis of perchloric acid extracts and identification of1H,31P and13C resonances. Biochim. Biophys. Acta 805: 104–116.

    Google Scholar 

  • Fan M-Y, Lum Z-P, Fu X-W, Levesque L, Tai IT and Sun AM (1990) Reversal of diabetes in BB rats by transplantation of encapsulated pancreatic islets. Diabetes 39: 519–522.

    Google Scholar 

  • Fernandez EJ, Mancuso A, Murphy MK, Blanch HW and Clark DS (1990) Nuclear magnetic resonance methods for observing the intracellular environment of mammalian cells. Ann. NY Acad. Sci. 589: 458–475.

    Google Scholar 

  • Gillies RF, Galons J-P, McGovern KA, Scherer PG, Lien Y-H, Job C, Ratcliff R, Chapa F, Cerdan S and Dale BE (1993) Design and application of NRM-compatible bioreactor circuits for extended perfusion of high-density mammalian cell cultures. NMR Biomedicine 6: 95–104.

    Google Scholar 

  • Grampp GE, Sambanis A and Stephanopoulos GN (1992) Use of regulated secretion in protein production from animal cells: an overview. Adv. Biochem. Eng. Biotechnol. 46: 35–62.

    Google Scholar 

  • Jans AWH and Willem R (1988)13C-NMR study of glycerol metabolism in rabbit renal cells of proximal convoluted tubulus. Eur. J. Biochem. 174: 64–73.

    Google Scholar 

  • Lacy PE, HEgre OD, Gerasimidi-Vazeou A, Gentile FT and Dionne KE (1991) Maintenance of normoglycemia in diabetic mice by subcateneous xenografts of encapsulated islets. Science 254: 1782–1784.

    Google Scholar 

  • Lanza RP, Butler DH, Borland KM, Staruk JE, Faustman DL, Solomon BA, Muller TE, Rupp RG, Maki T, Monaco AP and Chick WL (1991) Xenctransplantation of canine, bovine, and procine islets in diabetic rats without immunosuppression. Proc. Natl.Acad. Sci USA 88: 11100–11104

    Google Scholar 

  • Levesque L, Brubaker PL and Sun AM (1992) Maintenance of longterm secretory function by microencapsulated islets of Langerhands. Endocrinology 130: 644–650.

    Google Scholar 

  • Lim F and Sun AM (1980) Microencapsulated islets as bioartificial endocrine pancreas. Science 210: 908–910.

    Google Scholar 

  • Minichiello MM, Albert DM, Kolodny NH, Lee M-N and Craft JL (1989) A perfusion system developed for31P NMR study of melanoma cells at tissue-like density. Magn. Res. Med. 10: 96–107.

    Google Scholar 

  • Moon RB and Richards JEL (1973) Determination of intracellular pH by31P magnetic resonance. J. Biol. Chem. 248: 7276–7278.

    Google Scholar 

  • Moore H-PH, Walker MD, Lee F and Kelly RB (1983) Expressing a human proinsulin cDNA in a mouse ACTH-secreting cell. Intracellular storage, proteolytic processing, and secretion on stimulation. Cell 35: 531–538.

    Google Scholar 

  • Mosher TJ, Williams GD, Doumen C, LaNoue KF and Smith MB (1992) Error in the calibration of the MgATP chemical-shift limit: effects on the determination of free magnesium by31P NMR spectroscopy. Magn. Res. Med. 24: 163–169.

    Google Scholar 

  • Narayan KS, Mores EA, Chatham JC and Barker PB (1990)31P NMR of mammalian cells encapsulated in alginate gels utilizing a new phosphate-free perfusion medium. NMR Biomed. 3: 23–26.

    Google Scholar 

  • Neeman M and Degani H (1989) Early estrogen-induced metabolic changes and their inhibition by actinomycin D and cycloheximide in human breast cancer cells:31P and13C NMR studies. Proc. Natl. Acad. Sci. USA 86: 5585–5589.

    Google Scholar 

  • Papas KK (1992) Characterization of the metabolic and secretory behavior of suspended free and entrapped AtT-20 spheroids in fed-batch and perfusion cultures. MS thesis, Georgia Institute of Technology, Atlanta GA.

    Google Scholar 

  • Papas KK, Constantinidis I and Sambanis A (1993) Cultivation of recombinant, insulin-secreting AtT-20 cells as free and entrapped spheroids. Cytotechnology 13: 1–12.

    Google Scholar 

  • Peura RA (1988) Comparison of artificial pancreas devices. In: Skalak R and Fox CF (eds.). Tissue Engineering: Proceedings of a Workshop Held at Granlibakken, Lake Tahoe, California, February 26–29, pp. 223–230.

  • Ronen S and Degani H (1989) Studies of the metabolism of human breast cancer spheroids by NMR. Magn. Res. Med. 12: 274–281.

    Google Scholar 

  • Sambanis A, Papas KK, Constantinidis I and Long RC Jr (1993) Towards the development of a bioartificial pancreas: Fabrication and non-invasive monitoring of microbeads containing insulinsecreting, transformed cells. Presented at Annual Meeting of the American Institute of Chemical Engineers, St. Louis, Missouri, November 1993.

  • Sambanis A, Papas KK and Constantinidis I (1994) Towards the development of artificial endocrine tissues: I. Stability and function of immunoisolated, transformed insulin-secreting cells. Submitted.

  • Schnall MD, Subramanian VH, Leigh JS and Chance B (1985) A new double-tuned probe for concurrent1H and31P NMR. J. Magn. Reson. 65: 122–129.

    Google Scholar 

  • Seton MV (1989) Blood guts and chemical engineering. Can. J. Chem. Eng. 67: 705–712.

    Google Scholar 

  • Sherry AD, Malloy CR, Roby RE, Rajagopal A and Jeffrey FMH (1988) Propionate metabolism in the rat heart by13C NMR spectroscopy. Biochem. J. 254: 593–598.

    Google Scholar 

  • Soon-Shiong P, Feldman E, Nelson R, Komtebedde J, Smidsrod O, Skjak-Braek G, Espevik T, Heintz R and Lee M (1992) Successful reversal of spontaneous diabetes in dogs by intraperitoneal microencapsulated islets. Transplantation 54: 769–774.

    Google Scholar 

  • Soon-Shiong P, Lu ZN, Grewal I, Lanza RP and Clark W (1990) An in vitro method of assessing the immunoprotective properties of microcapsule membranes using pancreatic and tumor cell targets. Transplantation Proceedings 22: 754–755.

    Google Scholar 

  • Soon-Shiong P, Heintz RE, Merideth N, Yao QX, Yao Z, Zheng T, Murphy M, Moloney MK, Schmehl M, Harris M, Mendez R, Mendez R and Sandford PA (1994) Insulin independence in a type I diabetic patient after encapsulated islet transplantation. Lancet 343: 950–951.

    Google Scholar 

  • Stinson SC (1991) New drugs under development for diabetes. Chem. Eng. News, September 30: 35–59.

    Google Scholar 

  • Sullivan SJ, Maki T, Carretta M, Ozato H, Borland K, Mahoney MD, Muller TE, Solomon BA, Monaco AP and Chick WL (1992) Evaluation of the hybrid artificial pancreas in diabetic dogs. ASAIO Journal 38: 29–33.

    Google Scholar 

  • Sun AM (1988) Microencapsulation of pancreatic islet cells: a bicartificial endocrine pancreas. Methods Enzymol. 137: 575–580.

    Google Scholar 

  • Tal M, Thorens B, Surana M, Fleischer N, Lodish HF, Hanahan D and Efrat S (1992) Glucose transporter isotypes switch in T-antigen-transformed pancreatic β cells growing in culture and in mice. Mole. Cell. Biol. 12: 422–432.

    Google Scholar 

  • Tziampazis E (1993) Engineering functional, insulin-secreting cell systems: Effect of entrapment on cellular environment and secretory response. MS thesis, Georgia Institute of Technology, Atlanta GA.

    Google Scholar 

  • Tziampazis E and Sambanis A (1994) Tissue engineering a bioartificial pancreas: Modeling the cell envirnment and device function. Biotechnol. Progress, in press.

  • Vorlop K-D and Klein J (1983) New developments in the field of cell immobilization-formation of biocatalysis by ionotropic gelation. In: Lafferty RM (ed.) Enzyme Technology. III. Rotenburg Fermentation Symposium 1982, 22–24 September 1982, pp. 219–235. New York: Springer-Verlag.

    Google Scholar 

Download references

Author information

Authors and Affiliations

  1. School of Chemical Engineering, Georgia Institute of Technology, 30332-0100, Atlanta, GA, USA

    A. Sambanis & K. K. Papas

  2. Frederik Philips Magnetic Resonance Research Center, Department of Radiology, Emory University, School of Medicine, 30322, Atlanta, GA, USA

    P. C. Flanders, R. C. Long Jr., H. Kang & I. Constantinidis

Authors
  1. A. Sambanis
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. K. K. Papas
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. P. C. Flanders
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. R. C. Long Jr.
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. H. Kang
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. I. Constantinidis
    View author publications

    You can also search for this author in PubMed Google Scholar

Rights and permissions

Reprints and permissions

About this article

Cite this article

Sambanis, A., Papas, K.K., Flanders, P.C. et al. Towards the development of a bioartificial pancreas: Immunoisolation and NMR monitoring of mouse insulinomas. Cytotechnology 15, 351–363 (1994). https://doi.org/10.1007/BF00762410

Download citation

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF00762410

Key Words

Search

Navigation