Improved functional survival of human islets of langerhans in three-dimensional matrix culture

Mathias D. Brendel, Shen Shen Kong, Rodolfo Alejandro, Daniel H. Mintz

Research output: Contribution to journalArticle

51 Citations (Scopus)

Abstract

The current study evaluates functional survival of human islets maintained in tissue culture for up to 4 wk in suspension media (CMRL-1066 with supplements) and contrasts these results with immobilizing three-dimensional matrices (agarose or alginate). The absolute number and volume of islets retrieved from agarose is significantly higher after two and four wk of culture compared to conventional free-floating media. In vitro function of islets, assessed by insulin/DNA content, insulin secretion into the culture media over 24 h and glucose-theophylline stimulated insulin release in a dynamic perifusion system, was not significantly different between free-floating and matrix preserved islets. In vivo islet function was evaluated by the effectiveness for reversal of insulin-dependent diabetes mellitus by transplantation of the islets under the kidney capsule of nude mice. Although adequate insulin responses to glucose were seen after culture in conventional or matrix media, only agarose embedded islets were consistently able to induce normoglycemia in diabetic recipients after 14 days of culture. Additional transplantation experiments defined the threshold level required to reverse diabetes to be between 1,000 and 1,500 agarose preserved islets. Our data suggest improved engraftment of human islets after agarose culture. This culture method may be of benefit for the accumulation of functionally competent human islets, thus facilitating the implementation of clinical protocols that utilize freshly isolated islets from multiple donors without the need for cryopreservation.

Original languageEnglish
Pages (from-to)427-435
Number of pages9
JournalCell Transplantation
Volume3
Issue number5
StatePublished - Sep 1 1994

Fingerprint

Insulin
Islets of Langerhans
Sepharose
Medical problems
Glucose
Islets of Langerhans Transplantation
Tissue culture
Alginate
Cryopreservation
Theophylline
Clinical Protocols
Type 1 Diabetes Mellitus
Nude Mice
Capsules
Culture Media
Suspensions
Dynamical systems
DNA
Transplantation
Kidney

Keywords

  • Culture
  • Human islets of Langerhans
  • Immobilization
  • Transplantation

ASJC Scopus subject areas

  • Transplantation
  • Cell Biology

Cite this

Improved functional survival of human islets of langerhans in three-dimensional matrix culture. / Brendel, Mathias D.; Kong, Shen Shen; Alejandro, Rodolfo; Mintz, Daniel H.

In: Cell Transplantation, Vol. 3, No. 5, 01.09.1994, p. 427-435.

Research output: Contribution to journalArticle

Brendel, Mathias D. ; Kong, Shen Shen ; Alejandro, Rodolfo ; Mintz, Daniel H. / Improved functional survival of human islets of langerhans in three-dimensional matrix culture. In: Cell Transplantation. 1994 ; Vol. 3, No. 5. pp. 427-435.
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