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|Title:||Derivation of functional insulin-producing cell lines from primary mouse embryo culture|
|Authors:||Li, G.D. |
Way, Tan E.K.
|Citation:||Li, G.D., Luo, R., Zhang, J., Xie, F., Lim, S.K., Salto-Tellez, M., Yeo, K.S., Way, Tan E.K., Caille, D., Meda, P., Que, J., Lim, S.K., Kon, O.L. (2009). Derivation of functional insulin-producing cell lines from primary mouse embryo culture. Stem Cell Research 2 (1) : 29-40. ScholarBank@NUS Repository.|
|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.|
|Source Title:||Stem Cell Research|
|Appears in Collections:||Staff Publications|
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