Development of a novel β-cell specific promoter system for the identification of insulin-producing cells in in vitro cell cultures

Takuya Fukazawa, Junji Matsuoka, Yoshio Naomoto, Toru Nakai, Mary L. Durbin, Itaru Kojima, Jonathan R.T. Lakey, Noriaki Tanaka

Research output: Contribution to journalArticle

11 Citations (Scopus)

Abstract

Recently, it has been reported that islet transplantation into patients with Type 1 diabetes may achieve insulin independence for a year or longer [Shapiro et al., Islet transplantation in seven patients with type 1 diabetes mellitus using a glucocorticoid-free immunosuppressive regimen, N Engl J Med. 343 (2000) 230-238]. However, the amount of donor islet tissue is limited, therefore, multiple approaches are being explored to generate insulin-producing cells in vitro. Some promising results have been obtained using mouse and human stem cells and progenitor cells [Soria et al., From stem cells to beta cells: new strategies in cell therapy of diabetes mellitus, Diabetologia. 4 (2001) 407-415; Lechner et al., Stem/progenitor cells derived from adult tissues: potential for the treatment of diabetes mellitus, Am J Physiol Endocrinol Metab. 284 (2003) 259-266; Bonner-Weir et al., In vitro cultivation of human islets from expanded ductal tissue, Proc Natl Acad Sci U S A, 97 (2000) 7999-8004; Assady et al., Insulin production by human embryonic stem cells, 50 (2001) Diabetes 1691-1697]. However, the efficiency of obtaining populations with high numbers of differentiated cells has been poor. In order to improve the efficiency of producing and selecting insulin-producing cells from undifferentiated cells, we have designed a novel β-cell specific and glucose responsive promoter system designated pGL3.hINS-363 3×. This artificial promoter system exhibits significant luciferase activity not only in insulin-producing MIN6 m9 cells but also in isolated human islets. The pGL3.hINS-363 3× construct shows no activity in non-insulin-producing cells in low glucose conditions (2 mM glucose) but demonstrates significant activity and β-cell specificity in high glucose conditions (16 mM glucose). Furthermore, pGL3.hINS-363 3× shows significant promoter activity in differentiated AR42J cells that can produce insulin after activin A and betacellulin treatment. Here, we describe a novel β-cell specific and glucose responsive artificial promoter system designed for analyzing and sorting β-like insulin-producing cells that have differentiated from stem cells or other progenitor cells.

Original languageEnglish
Pages (from-to)3404-3412
Number of pages9
JournalExperimental Cell Research
Volume312
Issue number17
DOIs
Publication statusPublished - Oct 15 2006

Keywords

  • Diabetes mellitus
  • Differentiation
  • Insulin promoter
  • Islet transplantation
  • Stem cell
  • Transcription factor
  • β-cell

ASJC Scopus subject areas

  • Cell Biology

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