Derivation of functional insulin-producing cell lines from primary mouse embryo culture |
| |
Authors: | Guo Dong Li Ruihua Luo Jiping Zhang Keng Suan Yeo Fei Xie Eileen Khia Way Tan Dorothée Caille Jianwen Que Oi Lian Kon Manuel Salto-Tellez Paolo Meda Sai Kiang Lim |
| |
Affiliation: | 1. Cardiovascular Research Institute, National University Medical Institutes, Yong Loo Lin School of Medicine, National University of Singapore, Singapore 117597;2. Genome Institute of Singapore, ASTAR, Singapore;3. Department of Cell Physiology and Metabolism, University of Geneva, Geneva, Switzerland;4. Institute of Medical Biology, ASTAR, Singapore 138648;5. National Cancer Center, Singapore;6. Department of Pathology, National University of Singapore, Singapore 117597;7. Department of Surgery, Yong Loo Lin School of Medicine, National University of Singapore, Singapore 117597;8. School of Biological Sciences, Nanyang Technological University, Singapore |
| |
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. |
| |
Keywords: | |
本文献已被 ScienceDirect 等数据库收录! |
|