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Labelling of human adipose-derived stem cells for non-invasive in vivo cell tracking

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Abstract

Human adipose-derived stem cells (ASC) can be expanded in an undifferentiated state or differentiated along the osteogenic, chondrogenic, adipogenic, myogenic, endothelial and neurogenic lineage. To test their in vivo and in situ regenerative potential, their fate needs to be traced after application in suitable defect models. Non-invasive imaging systems allow for real time tracking of labelled cells in the living animal. We have evaluated a bioluminescence cell tracking approach to visualise ASC labelled with luciferase in the living animal. Two procedures have been tested to efficiently label human stem cells with a reporter gene (luciferase, green fluorescent protein), namely lipofection with LipofectamineTM 2000 and electroporation with a Nucleofector® device. With both lipofection and nucleofection protocols, we have reached transfection efficiencies up to 60%. Reporter gene expression was detectable for 3 weeks in vitro and did not interfere with the phenotype and the stem cell properties of the cells. By means of a highly sensitive CCD camera, we were able to achieve real time imaging of cell fate for at least 20 days after application (intravenous, intramuscular, intraperitoneal, subcutaneous) in nude mice. Moreover, we were able to influence cell mobility by choosing different modes of application such as enclosure in fibrin matrix. The optical imaging system with transient transfection is an elegant cell-tracking concept to follow survival and fate of human stem cells in small animals.

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Abbreviations

AP:

Alkaline Phosphatase

ASC:

Adipose-derived stem cells

BSA:

Bovine serum albumin

CCD:

Charge-coupled device

DAPI:

4,6-Diamidino-2-phenylindole

DMEM:

Dulbecco’s minimum essential medium

EDTA:

Ethylenediaminetetraacetic acid

EGM-2:

Endothelial growth medium-2

FCS:

Foetal calf serum

FISH:

Fluorescence in situ hybridisation

GFP:

Green fluorescent protein

HEPES:

4-2-Hydroxyethyl-1-piperazineethanesulfonic acid

i.m.:

Intramuscular

i.p.:

Intraperitoneal

i.v.:

Intravenous

MSC:

Mesenchymal stem cells

NOD/SCID:

Non-obese diabetic/severe combined immunodeficiency

PBS:

Phosphate buffered saline

SC:

Stem cells

s.c.:

Subcutaneous

SSC:

Saline, sodium citrate

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Acknowledgements

This work was partially supported by the European STREP Project HIPPOCRATES (NMP3-CT-2003-505758) and the Lorenz Boehler Fonds and was carried out under the scope of the European NoE EXPERTISSUES (NMP3-CT-2004-500283). We wish to thank Mika Brejnikow, Daniela Dopler, Kerstin Schauer and Tatjana Sindelar for excellent technical assistance.

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Authors and Affiliations

  1. Red Cross Blood Transfusion Service of Upper Austria, Blumauerstr. 3-5, Linz, A-4020, Austria

    Susanne Wolbank, Simone Hennerbichler & Christian Gabriel

  2. Ludwig Boltzmann Institute for Experimental and Clinical Traumatology/AUVA Research Centre, Linz/Vienna, Austria

    Susanne Wolbank, Anja Peterbauer, Esther Wassermann, Simone Hennerbichler, Martijn van Griensven & Heinz Redl

  3. Bio-Products & Bio-Engineering AG, Vienna, Austria

    Simone Hennerbichler

  4. Institute for Applied Microbiology, University of Natural Resources and Applied Life Sciences, Vienna, Austria

    Regina Voglauer

  5. Department of Human Genetics – General Women’s and Children’s Hospital, Linz, Austria

    Hans-Christoph Duba

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  1. Susanne Wolbank
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  2. Anja Peterbauer
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  3. Esther Wassermann
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  4. Simone Hennerbichler
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  5. Regina Voglauer
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  6. Martijn van Griensven
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  7. Hans-Christoph Duba
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  8. Christian Gabriel
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  9. Heinz Redl
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Correspondence to Susanne Wolbank.

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Wolbank, S., Peterbauer, A., Wassermann, E. et al. Labelling of human adipose-derived stem cells for non-invasive in vivo cell tracking. Cell Tissue Banking 8, 163–177 (2007). https://doi.org/10.1007/s10561-006-9027-7

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  • DOI: https://doi.org/10.1007/s10561-006-9027-7

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