Derivation of functional insulin-producing cell lines from primary mouse embryo culture

7Citations
Citations of this article
17Readers
Mendeley users who have this article in their library.

Abstract

We have previously described the derivation of insulin-producing cell lines from mouse embryonic stem cells (mESCs) by differentiation of an intermediate lineage-restricted E-RoSH cell line through nutrient depletion in the presence of nicotinamide followed by limiting dilution. Here we investigated whether insulin-producing cell lines could be similarly derived directly from mouse embryo cells or tissues. Using a similar approach, we generated the RoSH2.K and MEPI-1 to -14 insulin-producing cell lines from the 5.5-dpc embryo-derived E-RoSH-analogous RoSH2 cell line and a 6.0-dpc mouse embryo culture, respectively. Insulin content was ∼8 μg/106 MEPI-1 cells and 0.5 μg/106 RoSH2.K cells. Like insulin-producing mESC-derived ERoSHK cell lines, both MEPI and RoSH2.K lines were amenable to repeated cycles of freeze and thaw, replicated for months with a doubling time of 3-4 days, and exhibited genomic, structural, biochemical, and pharmacological properties of pancreatic β-cells, including storage and release of insulin and C-peptide in an equimolar ratio. Transplantation of these cells also reversed hyperglycemia in streptozotocin-treated SCID mice and did not induce teratoma. Like ERoSHK cells, both RoSH2.K and MEPI-1 cells also induced hypoglycemia in the mice. Therefore, our protocol is robust and could reproducibly generate insulin-producing cell lines from different embryonic cell sources. © 2008 Elsevier B.V. All rights reserved.

Cite

CITATION STYLE

APA

Li, G. D., Luo, R., Zhang, J., Yeo, K. S., Xie, F., Way Tan, E. K., … Lim, S. K. (2009). Derivation of functional insulin-producing cell lines from primary mouse embryo culture. Stem Cell Research, 2(1), 29–40. https://doi.org/10.1016/j.scr.2008.07.004

Register to see more suggestions

Mendeley helps you to discover research relevant for your work.

Already have an account?

Save time finding and organizing research with Mendeley

Sign up for free