원문정보
초록
영어
Background: Pluripotent stem cells (PSCs) including embryonic stem cells (ESCs) and induced pluripotent stem cells (iPSCs) offer the immense therapeutic potential in stem cell-based therapy of degenerative disorders. However, clinical trials of human ESCs cause heavy ethical concerns. With the derivation of iPSCs established by reprogramming from adult somatic cells through the transgenic expression of transcription factors, this problems would be able to overcome. In the present study, we tried to differentiate porcine iPSCs (piPSCs) into endothelial cells (ECs) for stem cell-based therapy of vascular diseases. Methods: piPSCs (OSKMNL) were induced to differentiation into ECs in four differentiation media (APEL-2, APEL-2 + 50 ng/mL of VEGF, EBM-2, EBM-2 + 50 ng/ mL of VEGF) on cultured plates coated with matrigel® (1:40 dilution with DMEM/F-12 medium) for 8 days. Differentiation efficiency of these cells were exanimated using qRT-PCR, Immunocytochemistry, Western blotting and FACS. Results: As results, expressions of pluripotency-associated markers (OCT-3/4, SOX2 and NANOG) were higher observed in all porcine differentiated cells derived from piPSCs (OSKMNL) cultured in four differentiation media than piPSCs as the control, whereas endothelial-associated marker (CD-31) in the differentiated cells was not expressed. Conclusions: It can be seen that piPSCs (OSKMNL) were not suitable to differentiate into ECs in the four differentiation media unlike porcine epiblast stem cells (pEpiSCs). Therefore, it would be required to establish a suitable PSCs for differentiating into ECs for the treatment of cardiovascular diseases.
목차
INTRODUCTION
MATERIALS AND METHODS
Induction of porcine induced pluripotent stem cells (piPSCs)
Isolation and culture of swine umbilical vein endothelial cells (SUVECs)
In vitro differentiation of piPSCs into endothelial cells
Alkaline phosphatase (AP) activity
Immunocytochemistry
Quantitative real-time PCR (qRT-PCR)
Western blotting
Flow cytometry (FACS) analysis
Statistical analysis
RESULTS
Morphology of piPSCs cultured for differentiation of endothelial cells
Expressions of pluripotency-associated genes and endothelial-associated gene in differentiated cells derived from piPSCs in differentiation media
Immunocytochemistry of pluripotency-associated markers in differentiated cells derived from piPSCs in differentiation media
Expression of endothelial-associated marker (CD-31) in differentiated cells derived from piPSCs in differentiation media
Flow cytometry of endothelial-associated marker(CD-31) in differentiated cells derived from piPSCs in differentiation media
DISCUSSION
CONCLUSION
REFERENCES