Effects of iron oxide incorporation for long term cell tracking on MSC differentiation in vitro and in vivo

doi:10.1016/j.bbrc.2008.02.159

Biochemical and Biophysical Research Communications

Volume 369, Issue 4, 16 May 2008, Pages 1076-1081

https://doi.org/10.1016/j.bbrc.2008.02.159 Get rights and content

Abstract

Successful cell therapy will depend on the ability to monitor transplanted cells. With cell labeling, it is important to demonstrate efficient long term labeling without deleterious effects on cell phenotype and differentiation capacity. We demonstrate long term (7 weeks) retention of superparamagnetic iron oxide particles (SPIO) by mesenchymal stem cells (MSCs) in vivo, detectable by MRI. In vitro, multilineage differentiation (osteogenic, chondrogenic and adipogenic) was demonstrated by histological evaluation and molecular analysis in SPIO labeled and unlabeled cells. Gene expression levels were comaparable to unlabeled controls in adipogenic and chondrogenic conditions however not in the osteogenic condition. MSCs seeded into a scaffold for 21 days and implanted subcutaneously into nude mice for 4 weeks, showed profoundly altered phenotypes in SPIO labeled samples compared to implanted unlabeled control scaffolds, indicating chondrogenic differentiation. This study demonstrates long term MSC traceability using SPIO and MRI, uninhibited multilineage MSC differentiation following SPIO labeling, though with subtle but significant phenotypical alterations.

Section snippets

Experimental protocol

Cell culture. Human bone marrow stromal cells (BMSCs) were isolated from femoral shaft biopsies of a patient undergoing total hip replacement after written consent (MEC-2004-142). BMSCs were isolated from the aspirated marrow according to previously described procedures [14]. Cells from the fourth passage were used for the experiments. Mesenchymal stem cells purchased from Cambrex (Verviers, Belgium) were used in the in vivo study.

Cell labeling. Cells were labeled with Endorem (Guerbet S.A.,

Multilineage differentiation of adult human MSCs

To determine the multilineage potential of labeled cells, SPIO labeled and unlabeled MSCs were cultured either in control medium or medium supplemented with adipogenic, chondrogenic or osteogenic factors for 13 days. Lineage specific histological staining was then performed. For both labeled and unlabeled MSCs, 13 days culture in appropriate differentiation medium led to positive staining for adipogenic (Fig. 1C and D), chondrogenic (Fig. 1H and I) and osteogenic (Fig. 1M and N), demonstrated

Adipogenesis

Following culture with adipogenic factors, significantly elevated levels of fatty acid binding protein-4 (FABP4), also known as aP2, were observed compared to unlabeled, untreated control cultures for both labeled and unlabeled cells (Fig. 2A; P = 0.0001, Two way ANOVA with repeated measures, n = 3). SPIO labeling had no effect on this expression. The same effect was observed with peroxisome proliferator-acvitated receptor γ (PPARγ), another marker of adipogenesis. Fig. 4B shows a significant

Discussion

Several groups have demonstrated the differentiation potential of stem cells or progenitor cells following SPIO labeling using different transfection methods [19], [20], [21], [22]. However, these papers have not investigated chondrogenic differentiation, nor have they looked beyond basic histological/immunohistological analyses or at multilineage MSC differentiation. A recent paper by Heymer et al. performed gene expression analyses but did not examine the effects of in vivo implantation on

Acknowledgments

We thank Nicole Kops for her assistance with the histological staining of implanted scaffolds, Corina de Ridder for her assistance with the implantation of the collagen-GAG scaffolds and Gavin Houston, Applied Science Laboratory, GE Healthcare for critical review of the manuscript and for helping with the optimization of data acquisition. We also acknowledge financial support from the Programme for Research in Third Level Institutions, The Science Foundation Ireland’s President of Ireland Young

References (25)

D.R. Elgort et al.
A review of technical advances in interventional magnetic resonance imaging
Acad. Radiol.
(2005)
F.P. Barry et al.
Mesenchymal stem cells: clinical applications and biological characterization
Int. J. Biochem. Cell Biol.
(2004)
S.W. Cho et al.
Engineering of volume-stable adipose tissues
Biomaterials
(2005)
H. Kim et al.
In vivo bone formation by human marrow stromal cells in biodegradable scaffolds that release dexamethasone and ascorbate-2-phosphate
Biochem. Biophys. Res. Commun.
(2005)
J.R. Mauney et al.
In vitro and in vivo evaluation of differentially demineralized cancellous bone scaffolds combined with human bone marrow stromal cells for tissue engineering
Biomaterials
(2005)
E.W. Mandl et al.
Fibroblast growth factor-2 in serum-free medium is a potent mitogen and reduces dedifferentiation of human ear chondrocytes in monolayer culture
Matrix Biol.
(2004)
E.J. Uitterlinden et al.
Glucosamine decreases expression of anabolic and catabolic genes in human osteoarthritic cartilage explants
Osteoarthritis Cartilage
(2006)
K.A. Hinds et al.
Highly efficient endosomal labeling of progenitor and stem cells with large magnetic particles allows magnetic resonance imaging of single cells
Blood
(2003)
A.S. Arbab et al.
Intracytoplasmic tagging of cells with ferumoxides and transfection agent for cellular magnetic resonance imaging after cell transplantation: methods and techniques
Transplantation
(2003)
Z. Zhang et al.
In vitro imaging of single living human umbilical vein endothelial cells with a clinical 3 0-T MRI scanner
Magma
(2005)

J.A. Frank et al.

Clinically applicable labeling of mammalian and stem cells by combining superparamagnetic iron oxides and transfection agents

Radiology

(2003)

J. Ringe et al.

Stem cells for regenerative medicine: advances in the engineering of tissues and organs

Naturwissenschaften

(2002)

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¹: Both authors have contributed equally.

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Effects of iron oxide incorporation for long term cell tracking on MSC differentiation in vitro and in vivo

Abstract

Section snippets

Experimental protocol

Multilineage differentiation of adult human MSCs

Adipogenesis

Discussion

Acknowledgments

Acad. Radiol.

Int. J. Biochem. Cell Biol.

Biomaterials

Biochem. Biophys. Res. Commun.

Biomaterials

Matrix Biol.

Osteoarthritis Cartilage

Blood

Intracytoplasmic tagging of cells with ferumoxides and transfection agent for cellular magnetic resonance imaging after cell transplantation: methods and techniques

Transplantation

In vitro imaging of single living human umbilical vein endothelial cells with a clinical 3 0-T MRI scanner

Magma

Clinically applicable labeling of mammalian and stem cells by combining superparamagnetic iron oxides and transfection agents

Radiology

Stem cells for regenerative medicine: advances in the engineering of tissues and organs

Naturwissenschaften