Abstract
We set out to assess the feasibility of exploiting expression of the mCherry gene, after lentiviral infection, in order visualise bone marrow-derived human mesenchymal stem cells (hMSCs) by optical imaging, and to provide proof of principle of this approach as a method for cell tracking and quantification in pre-clinical models. Commercial hMSCs were infected with a lentiviral vector carrying the mCherry gene under the control of the phosphoglycerate kinase promoter. After extensive in vitro culture, infected hMSCs were analysed for viability, morphology, differentiation capability, and maintenance of fluorescence. Thereafter, mCherry-positive cells were transplanted into unilaterally 6-hydroxy-dopamine lesioned rats (an experimental model of Parkinson's disease). Our analysis showed that hMSCs can be efficiently transduced with the lentiviral vector, retaining their biological features even in the long term. Intrastriatally transplanted mCherry-positive hMSCs can be detected ex vivo by a sensitive cooled CCD camera, both in the whole brain and in serial slices, and relatively quantified. Our protocol was found to be a reliable means of studying the viability of implanted hMSCs. mCherry labelling appears to be readily applicable in the post-transplantation tracking of stem cells and could favour the rapid development of new therapeutic targets for clinical treatments.
| Original language | English |
|---|---|
| Pages (from-to) | 114-125 |
| Number of pages | 12 |
| Journal | Archives Italiennes de Biologie |
| Volume | 151 |
| Issue number | 3 |
| Publication status | Published - 2013 |
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Keywords
- 6-OHDA
- Bioluminescence
- Parkinson's disease
ASJC Scopus subject areas
- Medicine(all)
- Cell Biology
- Physiology
Cite this
A reliable indirect cell-labelling protocol for optical imaging allows ex vivo visualisation of mesenchymal stem cells after ransplantation in rodent brain. / Diana, V.; Libani, I. V.; Armentero, M. T.; Blandini, F.; Lucignani, G.; Silani, V.; Cova, L.; Ottobrini, L.
In: Archives Italiennes de Biologie, Vol. 151, No. 3, 2013, p. 114-125.Research output: Contribution to journal › Article
}
TY - JOUR
T1 - A reliable indirect cell-labelling protocol for optical imaging allows ex vivo visualisation of mesenchymal stem cells after ransplantation in rodent brain
AU - Diana, V.
AU - Libani, I. V.
AU - Armentero, M. T.
AU - Blandini, F.
AU - Lucignani, G.
AU - Silani, V.
AU - Cova, L.
AU - Ottobrini, L.
PY - 2013
Y1 - 2013
N2 - We set out to assess the feasibility of exploiting expression of the mCherry gene, after lentiviral infection, in order visualise bone marrow-derived human mesenchymal stem cells (hMSCs) by optical imaging, and to provide proof of principle of this approach as a method for cell tracking and quantification in pre-clinical models. Commercial hMSCs were infected with a lentiviral vector carrying the mCherry gene under the control of the phosphoglycerate kinase promoter. After extensive in vitro culture, infected hMSCs were analysed for viability, morphology, differentiation capability, and maintenance of fluorescence. Thereafter, mCherry-positive cells were transplanted into unilaterally 6-hydroxy-dopamine lesioned rats (an experimental model of Parkinson's disease). Our analysis showed that hMSCs can be efficiently transduced with the lentiviral vector, retaining their biological features even in the long term. Intrastriatally transplanted mCherry-positive hMSCs can be detected ex vivo by a sensitive cooled CCD camera, both in the whole brain and in serial slices, and relatively quantified. Our protocol was found to be a reliable means of studying the viability of implanted hMSCs. mCherry labelling appears to be readily applicable in the post-transplantation tracking of stem cells and could favour the rapid development of new therapeutic targets for clinical treatments.
AB - We set out to assess the feasibility of exploiting expression of the mCherry gene, after lentiviral infection, in order visualise bone marrow-derived human mesenchymal stem cells (hMSCs) by optical imaging, and to provide proof of principle of this approach as a method for cell tracking and quantification in pre-clinical models. Commercial hMSCs were infected with a lentiviral vector carrying the mCherry gene under the control of the phosphoglycerate kinase promoter. After extensive in vitro culture, infected hMSCs were analysed for viability, morphology, differentiation capability, and maintenance of fluorescence. Thereafter, mCherry-positive cells were transplanted into unilaterally 6-hydroxy-dopamine lesioned rats (an experimental model of Parkinson's disease). Our analysis showed that hMSCs can be efficiently transduced with the lentiviral vector, retaining their biological features even in the long term. Intrastriatally transplanted mCherry-positive hMSCs can be detected ex vivo by a sensitive cooled CCD camera, both in the whole brain and in serial slices, and relatively quantified. Our protocol was found to be a reliable means of studying the viability of implanted hMSCs. mCherry labelling appears to be readily applicable in the post-transplantation tracking of stem cells and could favour the rapid development of new therapeutic targets for clinical treatments.
KW - 6-OHDA
KW - Bioluminescence
KW - Parkinson's disease
UR - http://www.scopus.com/inward/record.url?scp=84897780949&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=84897780949&partnerID=8YFLogxK
M3 - Article
VL - 151
SP - 114
EP - 125
JO - Archives Italiennes de Biologie
JF - Archives Italiennes de Biologie
SN - 0003-9829
IS - 3
ER -