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[SPIO-labeled rat bone marrow mesenchymal stem cells: alterations of biological activity and labeling efficiency
Abstract:
This study aimed to characterize and magnetic resonance imaging (MRI) track the mesenchymal stem cells labeled with polylysine-coated superparamagnetic iron oxide (PLL-SPIO). Rat bone marrow derived mesenchymal stem cells (rMSCs) were labeled with 25, 50 and 100 microg/mL PLL-SPIO for 24 hours. The labeling efficiency was assessed by iron content, Prussian blue staining, electron microscopy and in vitro MR imaging. The labeled cells were also analyzed for cytotoxicity and differentiation potential. Electron microscopic observations and Prussian blue staining revealed that 75% -100% of cells were labeled with iron particles. PLL-SPIO did not show any cytotoxicity up to 100 microg/mL concentration. Both 25 microg/mL and 50 microg/mL PLL-SPIO labeled stem cells did not exhibit any significant alterations in the adipo/osteo/chondrogenic differentiation potential compared to unlabeled control cells. The lower concentration of 25 microg/mL iron labeled cells emitted an obvious dark signal in T1W, T2WI and T2 * WI MR image. The novel PLL-SPIO enables to label and track rMSCs for in vitro MRI without cellular alteration. Therefore PLL-SPIO may potentially become a better MR contrast agent especially in tracking the transplanted stem cells and other cells without compromising cell functional quality.
Insights
Polylysine-coated superparamagnetic iron oxide (PLL-SPIO) effectively labels rat mesenchymal stem cells for MRI tracking. The labeled cells maintained their differentiation potential and showed no cytotoxicity, indicating PLL-SPIO as a promising MRI contrast agent.
Area of Science:
- Biomedical Engineering
- Cell Biology
- Radiology
Background:
- Tracking transplanted cells non-invasively is crucial for regenerative medicine.
- Superparamagnetic iron oxide nanoparticles (SPIONs) are potential contrast agents for Magnetic Resonance Imaging (MRI).
- Polylysine coating can enhance SPIONs' biocompatibility and cellular uptake.
Purpose of the Study:
- To characterize and evaluate polylysine-coated superparamagnetic iron oxide (PLL-SPIO) for labeling and tracking rat mesenchymal stem cells (rMSCs) using MRI.
- To assess the impact of PLL-SPIO labeling on rMSC cytotoxicity and differentiation potential.
- To determine the optimal concentration of PLL-SPIO for effective in vitro MRI tracking.
Main Methods:
- Rat bone marrow derived mesenchymal stem cells (rMSCs) were labeled with varying concentrations of PLL-SPIO (25, 50, 100 microg/mL).
- Labeling efficiency was assessed via iron content, Prussian blue staining, and electron microscopy.
- Cytotoxicity and in vitro differentiation (adipo/osteo/chondrogenic) potential were evaluated.
- In vitro MRI was performed on labeled cells using T1W, T2WI, and T2*WI sequences.
Main Results:
- Electron microscopy and Prussian blue staining confirmed 75%-100% labeling efficiency.
- PLL-SPIO exhibited no cytotoxicity up to 100 microg/mL.
- rMSCs labeled with 25 and 50 microg/mL PLL-SPIO showed no significant alterations in differentiation potential.
- A concentration of 25 microg/mL PLL-SPIO produced a clear dark signal in all MRI sequences.
Conclusions:
- Novel PLL-SPIO effectively labels rMSCs for in vitro MRI tracking without compromising cell viability or function.
- PLL-SPIO demonstrates potential as an MRI contrast agent for tracking transplanted cells.
- This method offers a promising approach for monitoring cell therapies non-invasively.
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