چاپ صفحه |  صفحه نخست سامانه |  نویسندگان |  اطلاعات تفضیلی |  دانلود مقاله | دانشگاه علوم پزشکی تبریز |
| نویسنده | نفر چندم مقاله |
|---|---|
| فرزانه فتحی | اول |
| محمدرضا رشیدی شاهگلی | دوم |
| رضا رهبرقاضی | سوم |
| عنوان | متن |
|---|---|
| کلمات کلیدی | Human amniotic mesenchymal stem cells; SPR Biosensor; VEcadherin |
| خلاصه مقاله | Background and Aim: The mesenchymal-to-endothelial transition is an essential phenomenon during tissue regeneration, leading to an increased vascular density in target tissues. Thus, the development of highly sensitive and accurate biosensor approaches for detecting stem cells differentiation, particularly in the earliest steps, is more crucial than ever. In the current study, a label-free SPR (surface plasmon resonance) method was developed for the monitoring of the early-stage differentiation of mesenchymal stem cells into endothelial-like phenotype in term of VEcadherin over a period of 14 days. Methods: Human amniotic mesenchymal stem cells (hAMSCs) line (Cat No: C10680) was obtained from Iranian Biological Resource Center. Low glucose-DMEM (DMEM/LG, Gibco) cell media were used for the cultivation of hAMSCs. To induce endothelial differentiation, hAMSCs were maintained in endothelial cell growth media, M-199, supplemented with an EGM-2 cocktail (Cat No: C-22010, Promocell) and 2% fetal calf serum (FCS, Promocell) for 14 days. The medium was replenished every 2 to 3 days. The differentiation of hAMSCs into endothelial-like phenotype was investigated by SPR biosensor method and also with flow cytometry analysis and immunofluorescence microscopy on days 1, 2, 3, 5, 7 and 14. The morphological changes in relation to the endothelial acquisition were monitored through the experiment. A multi-parameter SPR device (MP-SPR Navi 210A, BioNavis Ltd, Tampere, Finland) with gold chips (BioNavis Ltd, Finland) was used to examine antibody-cell affinity interactions. Results: The combination of SPR method and Ab immobilization on-chip resulted in early detection of VE-cadherin as a cell surface marker in the endothelial differentiation of hAMSCs. Consistent with endothelial differentiation of SCs, a high level of VE-cadherin was encoded on the cell surface resulting in an increased attachment of the cells to the immobilized antibody on the chip surface. This attachment, in turn, caused profound changes in the refractive index values in the detection region which were reflected as increased RU intensities during differentiation stages. After subtracting the nonspecific binding responses, we recorded 0, 80, 120, 360, 510 and 610 RUs×10-4 for differentiating hMSCs following 1, 2, 3, 5, 7 and 14 days, respectively. The flow cytometric method was unable to discriminate the hAMSCs expressing VE-cadherin during the first 4 days of the differentiation, especially on day 0, 1 and 2, toward endothelial lineage. Conclusion: In the present study, SPR technique could sense the early stage differentiation of hAMSCs on day 3 in real-time and label-free form without affecting cell viability, but flow cytometry and fluorescent microscopy methods were not able to detect the cell differentiation at the same time. This sensitive method presents hopeful views for monitoring and identification of rare and specific cell populations like tumor cells, cancer stem cells and etc. |
| نام فایل | تاریخ درج فایل | اندازه فایل | دانلود |
|---|---|---|---|
| IMG_20181228_234304.jpg | 1397/10/08 | 870927 | دانلود |
| Pages from Binder1.pdf | 1397/10/08 | 217604 | دانلود |
| PS-063.pptx | 1397/10/08 | 1065958 | دانلود |
