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Isolation of human amniotic mesenchymal cells and their differentiation potential into islet-like cells in vitro / 南方医科大学学报
Journal of Southern Medical University ; (12): 5-10, 2011.
Article in Chinese | WPRIM | ID: wpr-267683
ABSTRACT
<p><b>OBJECTIVE</b>To isolate human amniotic mesenchymal cells (hAMCs) and investigate their transdifferentiation ability into islet-like cells in vitro.</p><p><b>METHODS</b>Human amnion was treated with the trypsin/EDTA to remove the amniotic epithelial cells and then incubated with collagenase I and dispase at 37 degrees celsius; overnight. The cells were collected by centrifugation and identified for the expressions of vimentin and SSEA-4 using immunofluorescence assay and for CD29, CD90, CD34, and CD45 using flow cytometry. RT-PCR was performed to detect the expressions of ACTG2, ACTA2, MMP2, Cripto, Sox2, LEFTYA, nanog, and Oct-4 in the cells. The differentiation potential of the isolated cells into inslet-like cells was assessed after a 14-day induction with the inducing factors by RT-PCR and immunofluorescence assay.</p><p><b>RESULTS</b>The hAMCs were capable of in vitro proliferation and passaging for 10 passages while retaining the normal karyotype. The isolated cells were positive for staining of vimentin and SSEA-4 and negative for CD34 and CD45; the CD29 and CD90 cells accounted for (91.5∓9.93)% and (48.7∓9.47)% of the cells, respectively. The hAMCs expressed several pluripotency-related genes, including Cripto, Sox2, LEFTYA, nanog, and Oct-4. After induction, endocrine-related genes were expressed in the islet-like cells, including PDX1, ngn3, insulin and glucagon.</p><p><b>CONCLUSION</b>We have successfully established the method for isolating hAMCs, which possess the potential of differentiation into islet-like cells in vitro.</p>
Subject(s)
Full text: Available Index: WPRIM (Western Pacific) Main subject: Physiology / Cells, Cultured / Islets of Langerhans / Cell Culture Techniques / Cell Biology / Cell Transdifferentiation / Mesenchymal Stem Cells / Amnion / Methods Type of study: Prognostic study Limits: Female / Humans Language: Chinese Journal: Journal of Southern Medical University Year: 2011 Type: Article

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Full text: Available Index: WPRIM (Western Pacific) Main subject: Physiology / Cells, Cultured / Islets of Langerhans / Cell Culture Techniques / Cell Biology / Cell Transdifferentiation / Mesenchymal Stem Cells / Amnion / Methods Type of study: Prognostic study Limits: Female / Humans Language: Chinese Journal: Journal of Southern Medical University Year: 2011 Type: Article