A new experimental protocol for preferential differentiation of mouse embryonic stem cells into insulin-producing cells

Ortwin Naujok, Flavio Francini, Sally Picton, Anne Jörns, Sigurd Lenzen, Clifford J. Bailey

Research output: Contribution to journalArticle

Abstract

Mouse embiyonic stem (ES) cells have the potential to differentiate into insulin-producing cells, but efficient protocols for in vitro differentiation have not been established. Here we have developed a new optimized four-stage differentiation protocol and compared this with an established reference protocol. The new protocol minimized differentiation towards neuronal progeny, resulting in a population of insulin-producing cells with ß-cell characteristics but lacking neuronal features. The yield of glucagon and somatostatin cells was negligible. Crucial for this improved yield was the removal of a nestin selection step as well as removal of culture supplements that promote differentiation towards the neuronal lineage. Supplementation of the differentiation medium with insulin and fetal calf serum was beneficial for differentiation towards monohor-monal insulin-positive cells. After implantation into diabetic mice these insulin-producing cells produced a time-dependent improvement of the diabetic metabolic state, in contrast to cells differentiated according to the reference protocol. Using a spinner culture instead of an adherent culture of ES cells prevented the differentiation towards insulin-producing cells. Thus, prevention of cell attachment in a spinner culture represents a means to keep ES cells in an undifferentiated state and to inhibit differentiation. In conclusion, this study describes a new optimized four-stage protocol for differentiating ES cells to insulin-producing cells with minimal neuronal cell formation. Copyright © 2008 Cognizant Comm. Corp.
Original languageEnglish
Pages (from-to)1231-1242
Number of pages12
JournalCell Transplantation
Volume17
Issue number10-11
DOIs
Publication statusPublished - 2008

Fingerprint

Insulin
Stem cells
Cell culture
Stem Cells
Cells
Mouse Embryonic Stem Cells
Nestin
Somatostatin-Secreting Cells
Glucagon
Cell Differentiation

Keywords

  • embryonic stem cells
  • differentiation
  • insulin
  • diabetes

Cite this

Naujok, Ortwin ; Francini, Flavio ; Picton, Sally ; Jörns, Anne ; Lenzen, Sigurd ; Bailey, Clifford J. / A new experimental protocol for preferential differentiation of mouse embryonic stem cells into insulin-producing cells. In: Cell Transplantation. 2008 ; Vol. 17, No. 10-11. pp. 1231-1242.
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A new experimental protocol for preferential differentiation of mouse embryonic stem cells into insulin-producing cells. / Naujok, Ortwin; Francini, Flavio; Picton, Sally; Jörns, Anne; Lenzen, Sigurd; Bailey, Clifford J.

In: Cell Transplantation, Vol. 17, No. 10-11, 2008, p. 1231-1242.

Research output: Contribution to journalArticle

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