Contributions
Abstract: P676
Type: Poster
Abstract Category: Therapy - disease modifying - Neurorepair
Background: Mesenchymal stem cell (MSC) transplantation is currently under investigation as a potential treatment for MS. The ability to track MSCs in vivo after administration is an important step in developing this therapy. Recent development of the paramagnetic cell label, Cell Sense (CS), provides a promising way for this analysis to be performed. CS is an aqueous colloidal nanosuspension of a perfluorocarbon polymer with high content of the nonradioactive 19F isotope, which permits detection of labelled cells by MRI.
Objectives: To assess the feasibility of labeling MSCs with CS and evaluate the effect on viability, phenotype, and function.
Methods: Optimization of CS labeling into the clinical protocol for ex-vivo expansion of healthy normal donor bone marrow derived MSCs was performed with both fresh and cryopreserved MSCs. CS dose and labeling time matrices were implemented to refine the procedure. CS uptake by MSCs was assessed by nuclear magnetic resonance (NMR). The impact of CS labeling on viability was assessed by trypan blue staining; phenotype by flow cytometric analysis of cell surface markers; and function by assessing protein levels of thirteen key signaling molecules (with CellPrintTM), a sensitive and quantitative flow cytometric staining technique.
Results: Labeling of fresh and frozen MSCs using an optimal dose of 5 mg/ml CS were incubated for 24 hours. CS uptake was three times more efficient with fresh cells (6.46 x 1011 atoms/cell) compared to recently thawed MSCs (2.19 x 1011 atoms/cell) as detected by NMR. Viability was not affected by labeling with CS at the optimized dose and time combination. CS labeling had no effect on the expression of MSC-specific cell surface of markers (CD105, CD90, CD73). Expression levels of p-AKT(308), Musashi-2, ATG7, FoxP1, VEGF, and TGF-b were moderately changed. No effect was observed for HGF, b-catenin, MyD88, iNOS, p-p38, p-ERK1/2 and MMP-1.
Conclusions: These data indicate that labeling MSCs with CS does not significantly affect the viability or phenotype. Whether there are changes in cell function requires further investigation. These results suggest that cell tracking by MRI of MSCs following IV infusion in humans is feasible with the use of CS.
Disclosure:
Sarah M Planchon: Nothing to disclose.
Jane Reese-Koc: Nothing to disclose.
Brittney Hooper: Nothing to disclose.
David Kaplan is an owner of CellPrint Biotechnology, LLC.
Nicholas Kaye is an employee of CellPrint Biotechnology, LLC.
Brooke Helfer is an employee at Celsense, Inc, manufacturer of Cell Sense, the 19F imaging reagent used herein.
Charles F O"Hanlon, III is an employee and shareholder of Celsense, Inc., the manufacturer of Cell Sense, the 19F imaging reagent used herein.
Jeffrey A Cohen reports personal compensation for consulting for Genentech, Genzyme, Novartis, and Receptos; as a speaker for Teva; and as Co-Editor of Multiple Sclerosis Journal - Experimental, Translational and Clinical.
Abstract: P676
Type: Poster
Abstract Category: Therapy - disease modifying - Neurorepair
Background: Mesenchymal stem cell (MSC) transplantation is currently under investigation as a potential treatment for MS. The ability to track MSCs in vivo after administration is an important step in developing this therapy. Recent development of the paramagnetic cell label, Cell Sense (CS), provides a promising way for this analysis to be performed. CS is an aqueous colloidal nanosuspension of a perfluorocarbon polymer with high content of the nonradioactive 19F isotope, which permits detection of labelled cells by MRI.
Objectives: To assess the feasibility of labeling MSCs with CS and evaluate the effect on viability, phenotype, and function.
Methods: Optimization of CS labeling into the clinical protocol for ex-vivo expansion of healthy normal donor bone marrow derived MSCs was performed with both fresh and cryopreserved MSCs. CS dose and labeling time matrices were implemented to refine the procedure. CS uptake by MSCs was assessed by nuclear magnetic resonance (NMR). The impact of CS labeling on viability was assessed by trypan blue staining; phenotype by flow cytometric analysis of cell surface markers; and function by assessing protein levels of thirteen key signaling molecules (with CellPrintTM), a sensitive and quantitative flow cytometric staining technique.
Results: Labeling of fresh and frozen MSCs using an optimal dose of 5 mg/ml CS were incubated for 24 hours. CS uptake was three times more efficient with fresh cells (6.46 x 1011 atoms/cell) compared to recently thawed MSCs (2.19 x 1011 atoms/cell) as detected by NMR. Viability was not affected by labeling with CS at the optimized dose and time combination. CS labeling had no effect on the expression of MSC-specific cell surface of markers (CD105, CD90, CD73). Expression levels of p-AKT(308), Musashi-2, ATG7, FoxP1, VEGF, and TGF-b were moderately changed. No effect was observed for HGF, b-catenin, MyD88, iNOS, p-p38, p-ERK1/2 and MMP-1.
Conclusions: These data indicate that labeling MSCs with CS does not significantly affect the viability or phenotype. Whether there are changes in cell function requires further investigation. These results suggest that cell tracking by MRI of MSCs following IV infusion in humans is feasible with the use of CS.
Disclosure:
Sarah M Planchon: Nothing to disclose.
Jane Reese-Koc: Nothing to disclose.
Brittney Hooper: Nothing to disclose.
David Kaplan is an owner of CellPrint Biotechnology, LLC.
Nicholas Kaye is an employee of CellPrint Biotechnology, LLC.
Brooke Helfer is an employee at Celsense, Inc, manufacturer of Cell Sense, the 19F imaging reagent used herein.
Charles F O"Hanlon, III is an employee and shareholder of Celsense, Inc., the manufacturer of Cell Sense, the 19F imaging reagent used herein.
Jeffrey A Cohen reports personal compensation for consulting for Genentech, Genzyme, Novartis, and Receptos; as a speaker for Teva; and as Co-Editor of Multiple Sclerosis Journal - Experimental, Translational and Clinical.