Effect of Extracellular Matrix on Differentiation of Mouse Embryonic Stem Cells into Insulin-Producing Cells
Objective: Evaluation of extracellular matrics (ECMs) effect on differentiation ofembryonic stem cells (ESCs) to pancreatic β-cell.Materials and Methods: Mouse ESC line, Royan B1, was subjected to differentiationinto β-like cells in a three-step method: generation of embryoid bodies(EBs), spontaneou...
| Main Authors: | , , , , |
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| Format: | Article |
| Language: | English |
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Royan Institute (ACECR), Tehran
2009-01-01
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| Series: | Cell Journal |
| Subjects: | |
| Online Access: | http://celljournal.org/library/upload/article/Article%203.pdf |
| _version_ | 1819111587218194432 |
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| author | A. Farrokhi E. Elahi M.Pirouz H. Jafary H. Baharvand |
| author_facet | A. Farrokhi E. Elahi M.Pirouz H. Jafary H. Baharvand |
| author_sort | A. Farrokhi |
| collection | DOAJ |
| description | Objective: Evaluation of extracellular matrics (ECMs) effect on differentiation ofembryonic stem cells (ESCs) to pancreatic β-cell.Materials and Methods: Mouse ESC line, Royan B1, was subjected to differentiationinto β-like cells in a three-step method: generation of embryoid bodies(EBs), spontaneous differentiation and induction by Nicotinamide onto differentmatrices including poly L-ornithine/laminin, gelatin, and two different dilution ofmatrigel (1:30, 1:100) and control group (no ECM). At the final step, differentiatedcells were analyzed for expression of some pancreas-specific genes using "semiquantitativeRT-PCR ", for detection of insulin and C-peptide presence in cells using"immunocytochemistry" and for the evaluation of the amount of secretd insulinin response to glucose Using "insulin secretion assay".Results: The semi-quantitative RT-PCR analysis of differentiated cells on 1:30matrigel coated-plates showed consistent higher expression of β-cell specificmarkers including Insulin I, Insulin II, Slc2a2 in comparison to the other ECMs.The results of immunostainig for C-peptide showed no significant differences betweenthe experimental groups and finally insulin secretion assay revealed thatdifferentiated cells on 1:30 matrigel coated-plates secreted more insulin in responseto glucose in comparison to the other ECMs.Conclusion: Our results suggest that type of ECM may influence ESC differentiationinto insulin-secreting cells and 1:30 matrigel was more effective. However, thesuccess rate of differentiation needs further investigations using other appropriateECMs. |
| first_indexed | 2024-12-22T03:59:59Z |
| format | Article |
| id | doaj.art-6f4de29988a94c6d87c77c9be9882bc2 |
| institution | Directory Open Access Journal |
| issn | 2228-5806 2228-5814 |
| language | English |
| last_indexed | 2024-12-22T03:59:59Z |
| publishDate | 2009-01-01 |
| publisher | Royan Institute (ACECR), Tehran |
| record_format | Article |
| series | Cell Journal |
| spelling | doaj.art-6f4de29988a94c6d87c77c9be9882bc22022-12-21T18:39:46ZengRoyan Institute (ACECR), TehranCell Journal2228-58062228-58142009-01-01104250259Effect of Extracellular Matrix on Differentiation of Mouse Embryonic Stem Cells into Insulin-Producing CellsA. FarrokhiE. ElahiM.PirouzH. JafaryH. BaharvandObjective: Evaluation of extracellular matrics (ECMs) effect on differentiation ofembryonic stem cells (ESCs) to pancreatic β-cell.Materials and Methods: Mouse ESC line, Royan B1, was subjected to differentiationinto β-like cells in a three-step method: generation of embryoid bodies(EBs), spontaneous differentiation and induction by Nicotinamide onto differentmatrices including poly L-ornithine/laminin, gelatin, and two different dilution ofmatrigel (1:30, 1:100) and control group (no ECM). At the final step, differentiatedcells were analyzed for expression of some pancreas-specific genes using "semiquantitativeRT-PCR ", for detection of insulin and C-peptide presence in cells using"immunocytochemistry" and for the evaluation of the amount of secretd insulinin response to glucose Using "insulin secretion assay".Results: The semi-quantitative RT-PCR analysis of differentiated cells on 1:30matrigel coated-plates showed consistent higher expression of β-cell specificmarkers including Insulin I, Insulin II, Slc2a2 in comparison to the other ECMs.The results of immunostainig for C-peptide showed no significant differences betweenthe experimental groups and finally insulin secretion assay revealed thatdifferentiated cells on 1:30 matrigel coated-plates secreted more insulin in responseto glucose in comparison to the other ECMs.Conclusion: Our results suggest that type of ECM may influence ESC differentiationinto insulin-secreting cells and 1:30 matrigel was more effective. However, thesuccess rate of differentiation needs further investigations using other appropriateECMs.http://celljournal.org/library/upload/article/Article%203.pdfEmbryonic Stem CellDifferentiationβ-CellExtracellular Matrix |
| spellingShingle | A. Farrokhi E. Elahi M.Pirouz H. Jafary H. Baharvand Effect of Extracellular Matrix on Differentiation of Mouse Embryonic Stem Cells into Insulin-Producing Cells Cell Journal Embryonic Stem Cell Differentiation β-Cell Extracellular Matrix |
| title | Effect of Extracellular Matrix on Differentiation of Mouse Embryonic Stem Cells into Insulin-Producing Cells |
| title_full | Effect of Extracellular Matrix on Differentiation of Mouse Embryonic Stem Cells into Insulin-Producing Cells |
| title_fullStr | Effect of Extracellular Matrix on Differentiation of Mouse Embryonic Stem Cells into Insulin-Producing Cells |
| title_full_unstemmed | Effect of Extracellular Matrix on Differentiation of Mouse Embryonic Stem Cells into Insulin-Producing Cells |
| title_short | Effect of Extracellular Matrix on Differentiation of Mouse Embryonic Stem Cells into Insulin-Producing Cells |
| title_sort | effect of extracellular matrix on differentiation of mouse embryonic stem cells into insulin producing cells |
| topic | Embryonic Stem Cell Differentiation β-Cell Extracellular Matrix |
| url | http://celljournal.org/library/upload/article/Article%203.pdf |
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