Cell viability evaluation of transdifferentiated endothelial-like cells by quantitative electron-probe X-ray...

Area of Science:

• Biomaterials Science
• Cell Biology
• Regenerative Medicine

Background:

• Vascular tissue engineering requires robust endothelial cell viability for functional grafts.
• Human umbilical cord Wharton's jelly stem cells (HWJSC) offer a potential source for endothelial cells.
• Transdifferentiation is a promising strategy to generate endothelial-like cells (Tr-ELC) from stem cells.

Purpose of the Study:

• To evaluate the cell viability of Tr-ELC using electron probe X-ray microanalysis (EPXMA).
• To quantify intracellular ionic concentrations in Tr-ELC, HWJSC, and HUVEC.
• To assess the suitability of Tr-ELC for vascular tissue engineering.

Main Methods:

• Isolation and culture of HWJSC and human umbilical vein endothelial cells (HUVEC).
• Induction of transdifferentiation from HWJSC to Tr-ELC.
• EPXMA analysis of HUVEC, HWJSC, and Tr-ELC using a scanning electron microscope and EDAX DX-4 system.
• Quantification of intracellular ion content via the peak-to-local-background (P/B) ratio method.

Main Results:

• Tr-ELC exhibited a high K/Na ratio (9.41), indicating robust viability.
• Intracellular levels of chlorine, phosphorus, and magnesium were maintained in Tr-ELC compared to HWJSC.
• Tr-ELC showed increased calcium and sulfur levels, with calcium levels similar to HUVEC.
• EPXMA confirmed Tr-ELC possess a microanalytical profile comparable to endothelial cells.

Conclusions:

• Transdifferentiated endothelial-like cells (Tr-ELC) demonstrate high viability and possess a similar microanalytical profile to endothelial cells.
• HWJSC-derived Tr-ELC meet the viability and microanalytical requirements for vascular tissue engineering.
• EPXMA is a valuable tool for assessing cell viability in tissue engineering applications.