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Characterization of Blood Outgrowth Endothelial Cells (BOEC) from Porcine Peripheral Blood
Published on: January 6, 2022
[In vitro MR imaging of Fe(2)O(3)-PLL labelled rabbit peripheral blood endothelial progenitor cells]
Xiao-li Mai1, Gao-jun Teng, Zhan-long Ma
1Laboratory of Molecular Imaging, Department of Radiology, Zhong Da Hospital, Southeast University, Nanjing 210009, China.
Objective:
To perform in vitro magnetic resonance imaging on magnetic iron oxide (Fe(2)O(3)-PLL) labeled rabbit peripheral blood endothelial progenitor cells (EPCs).
Methods:
Fe(2)O(3) was incubated with PLL for 2 hours to form Fe(2)O(3)-PLL. Rabbit peripheral blood mononuclear cells (MNCs) were isolated and EPCs were selected by adherence method, expanded and incubated with Fe(2)O(3)-PLL. Intracellular iron was detected by Prussian blue stain and under electron microscope. MTT assay was used to evaluate cell survival and proliferation of Fe(2)O(3)-PLL labeled EPCs. Flow cytometry was used to analysis cell cycle and apoptosis. The cells underwent in vitro MR imaging with various sequences.
Results:
Iron-containing intracytoplasmatic vesicles could be observed clearly with Prussian blue staining and electron microscope observation. Survival, life cycle and apoptosis values obtained by MTT and flow cytometry analysis were similar among unlabelled EPCs and EPCs labeled with various concentrations Fe(2)O(3)-PLL. The signal intensity on MRI was significantly decreased in labeled cells compared with that in unlabeled cells. The percentage change in signal intensity (DeltaSI) was most significant on T(2)*WI and DeltaSI was significantly lower in cells labeled for 7 days than that labeled for 1 day.
Conclusions:
The rabbit peripheral blood EPCs can be labeled with Fe(2)O(3)-PLL without significant change in viability and proliferation. The labeled EPCs can be imaged with standard 1.5 T MR equipment. The degree of MR signal decreasing may indirectly reflect the cells count, growth state and division.
Insights
Magnetic iron oxide nanoparticles successfully labeled rabbit endothelial progenitor cells (EPCs) for in vitro MRI. Labeled EPCs maintained viability and proliferation, enabling cell tracking via MRI signal changes.
Area of Science:
- Biomedical Engineering
- Cell Biology
- Nanotechnology
Context:
- Endothelial progenitor cells (EPCs) are crucial for vascular repair.
- Non-invasive cell tracking is essential for monitoring therapeutic cell delivery.
- Iron oxide nanoparticles offer potential as contrast agents for Magnetic Resonance Imaging (MRI).
Purpose:
- To evaluate the efficacy of Fe(2)O(3)-PLL nanoparticle labeling of rabbit peripheral blood EPCs.
- To assess the impact of Fe(2)O(3)-PLL labeling on EPC viability, proliferation, and cell cycle.
- To determine the feasibility of in vitro MRI for detecting and quantifying labeled EPCs.
Summary:
- Rabbit peripheral blood EPCs were isolated and labeled with Fe(2)O(3)-PLL nanoparticles.
- Cell viability, proliferation, and cell cycle were unaffected by labeling, confirmed by MTT and flow cytometry.
- Prussian blue staining and electron microscopy confirmed intracellular iron uptake.
- In vitro MRI demonstrated significant signal reduction in labeled EPCs, particularly on T(2)*-weighted images.
- Signal intensity changes correlated with labeling duration, suggesting potential for cell quantification.
Impact:
- Fe(2)O(3)-PLL labeling is a safe and effective method for preparing EPCs for MRI.
- This technique allows for non-invasive monitoring of labeled EPCs using standard 1.5 T MRI scanners.
- MRI signal alterations may serve as an indirect indicator of cell number, growth status, and division in vitro.
