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>
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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