Mesenchymal Stem Cells: Isolation, Characterisation and In Vivo Fluorescent Dye Tracking

Weir, Christopher; Morel-Kopp, Marie-Christine; Gill, Anthony J.; Tinworth, Kellie D.; Ladd, Leigh A.; Hunyor, Stephen N.; Ward, Christopher

doi:10.1016/j.hlc.2008.01.006

Cited by 88 publications

(79 citation statements)

References 21 publications

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“…18 Eventually, we also evaluated a fourth family of strategies consisting of using an antibody labeling kit (IRDye 800CW) to covalently stain the extracellular domain of membrane proteins. As previous studies have nicely shown that carbocyanine lipophilic dyes are very effective tools for WBI and/or IVM tracking of normal and malignant bone marrow (BM)-derived cells without affecting their functionality, [19][20][21][22][23] we aimed at using similar strategies to track primary human acute myeloid leukemia (AML) and hematopoietic stem cells (HSCs).…”

Section: Introductionmentioning

confidence: 99%

“Microenvironmental contaminations” induced by fluorescent lipophilic dyes used for noninvasive in vitro and in vivo cell tracking

Lassailly

Griessinger

Bonnet

2010

Blood

114

100

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Determining how normal and leukemic stem cells behave in vivo, in a dynamic and noninvasive way, remains a major challenge. Most optical tracking technologies rely on the use of fluorescent or bioluminescent reporter genes, which need to be stably expressed in the cells of interest. Because gene transfer in primary leukemia samples represents a major risk to impair their capability to engraft in a xenogenic context, we evaluated the possibility to use gene transfer-free labeling technologies. The lipophilic dye 3,3,3,3 tetramethylindotricarbocyanine iodide (DiR) was selected among 4 nearinfrared (NIR) staining technologies. Unfortunately we report here a massive transfer of the dye occurring toward the neighbor cells both in vivo and in vitro. We further demonstrate that all lipophilic dyes tested in this study (1,1-dioctadecyl-3,3,3,3-tetramethylindotricarbocyanine perchlorate [DiI], DiD, DiR, and PKH26) can give rise to microenvironmental contamination, including when used in suboptimal concentration, after extensive washing procedures and in the absence of phagocytosis or marked cell death. This was observed from all cell types tested. Eventually, we show that this microenvironmental contamination is mediated by both direct cell-cell contacts and diffusible microparticles. We conclude that tracking of labeled cells using non-genetically encoded markers should always be accompanied by drastic cross validation using multimodality approaches. (Blood. 2010;115(26): 5347-5354) IntroductionDespite recent animated discussions, 1 xenotransplantation of human cells in immunodeficient animals remains the only available technique to analyze the "stemness" of human cancer-initiating cells. [2][3][4][5] Despite the wide use of this model, homing and dynamic behavior of injected cells depending on time remain largely unknown both for normal and malignant cells. Whole-body imaging (WBI) and intravital microscopy (IVM) provide powerful tools to longitudinally monitor the behavior of cancers in vivo 6,7 and to track and analyze normal and cancer-initiating cells in their microenvironment or niche. 8,9 Near-infrared optical imaging (NIR) represents a very attractive technology in this context by offering a relatively easy and cheap access to multimodality and multiscale in vivo imaging. 10,11 In vivo cell tracking using optical imaging technologies largely relies on the use of ectopic expression of reporter genes such as Luciferase (for bioluminescence) or on fluorescent proteins. To obtain such expression, cells have to be engineered in vitro so that the reporter gene can be integrated in their genome for stable expression over time. Because in vitro manipulation of primary leukemia samples represents a real risk to impede the functionality of the cells, gene transfer-free approaches would represent a major improvement in this context. 12 Various strategies have been reported to fluorescently label cells in vitro for subsequent tracking in vivo. [13][14][15][16] In the field of immunohematology, the succinimidyl ester of carb...

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Section: Introductionmentioning

confidence: 99%

“Microenvironmental contaminations” induced by fluorescent lipophilic dyes used for noninvasive in vitro and in vivo cell tracking

Lassailly

Griessinger

Bonnet

2010

Blood

114

100

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“…In the presence of Netrin-1, its receptors in colorectal cancer, (DCC) and UNC5H transduce signalling cascades that lead to the proliferation, migration and differentiation of the cells. 28 Using a Matrigel angiogenesis assay, we also discovered augmented angiogenesis and arteriogenesis (blood vessels ⩾20 µm in diameter) in the Netrin-1-BMSCs group compared to Null-BMSCs, while in the control group, virtually no angiogenesis was observed. New angiogenesis consisted of ECs, which were mostly derived from the soma, with only a few ECs differentiated from BMSCs.…”

Section: Discussionmentioning

confidence: 82%

“…Because BMSCs can secrete multiple angiogenic cytokines, differentiate into ECs and SMCs that incorporate into the vessels and be amplified in vitro to generate a considerable number of high-quality cells for transplantation 26 ; BMSC transplantation has been considered a fitting treatment for ischaemic diseases with the absence of immunosuppression and ethical problems. 27,28 Although therapeutic transplantation of BMSCs alone in ischaemic disease can achieve some improvements, the therapeutic effect is not marked in all patients and even failed in some. The major reason for failure is the low survival rate of transplanted cells.…”

Section: Discussionmentioning

confidence: 99%

Netrin-1 promotes mesenchymal stem cell revascularization of limb ischaemia

Liu

Wang

et al. 2016

Diabetes and Vascular Disease Research

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This study examines the effect and mechanism of action of Netrin-1 on bone marrow mesenchymal stem cells in angiogenesis. Tube formation and migration of bone marrow mesenchymal stem cells were observed in cell culture. Bone marrow mesenchymal stem cells or Netrin-1-bone marrow mesenchymal stem cells were injected into the ischaemic area of the rat hind limb on the first day after surgery. Laser Doppler perfusion imaging was performed to analyse the levels of vascular endothelial growth factor in plasma and muscles, and immunohistochemistry and immunofluorescence were used to analyse angiogenesis. Bone marrow mesenchymal stem cells in medium containing Netrin-1 markedly increased the number of tubes formed and the migration of bone marrow mesenchymal stem cells compared with the untreated control group. The function of Netrin-1 in tube formation and migration is similar to vascular endothelial growth factor, and combined with vascular endothelial growth factor, Netrin-1 has more enhanced effect than in the other three groups. The Netrin-1-bone marrow mesenchymal stem cell group had better augmented blood-perfusion scores and vessel densities, as well as improved function of the ischaemic limb than that of the group injected with bone marrow mesenchymal stem cells (treated with bone marrow mesenchymal stem cells individually) or the control group (treated with medium). These results suggest that Netrin-1 has the ability to augment the angiogenesis of bone marrow mesenchymal stem cells and improve the function of the ischaemic hind limb by increasing the level of vascular endothelial growth factor.

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“…Dog CSCs can also form three-dimensional cell aggregates, so called cardiospheres, whose regenerative properties are currently under investigation (Bartosh et al 2008). Sheep mesenchymal stem cells can be easily derived as in other large animal models (Weir et al 2008). In this species the perivascular STRO-3 positive mesenchymal precursor cells (MPCs), a small fraction of mesenchymal stem cells that demonstrate an extensive capacity for proliferation and differentiation, have also been identified and tested for their regenerative properties (Hamamoto et al 2009).…”

Section: Endothelial and Epicardial Progenitor Cellsmentioning

confidence: 99%

Isolation, Characterization and Differentiation Potential of Cardiac Progenitor Cells in Adult Pigs

Vanelli

Pennarossa

Maffei

et al. 2012

Stem Cell Rev and Rep

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Mesenchymal Stem Cells: Isolation, Characterisation and In Vivo Fluorescent Dye Tracking

Cited by 88 publications

References 21 publications

“Microenvironmental contaminations” induced by fluorescent lipophilic dyes used for noninvasive in vitro and in vivo cell tracking

“Microenvironmental contaminations” induced by fluorescent lipophilic dyes used for noninvasive in vitro and in vivo cell tracking

Netrin-1 promotes mesenchymal stem cell revascularization of limb ischaemia

Isolation, Characterization and Differentiation Potential of Cardiac Progenitor Cells in Adult Pigs

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