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Identification of a putative pathway for the muscle homing of stem cells in a muscular dystrophy model.

Torrente Y, Camirand G, Pisati F, Belicchi M, Rossi B, Colombo F, El Fahime M, Caron NJ, Issekutz AC, Constantin G, Tremblay JP, Bresolin N - J. Cell Biol. (2003)

Bottom Line: The subpopulation of Sca-1+/CD34- MDSCs expressing L-selectin was called homing MDSCs (HMDSCs).Importantly, we found that vascular endothelium from striate muscle of young mdx mice expresses mucosal addressin cell adhesion molecule-1 (MAdCAM-1), a ligand for L-selectin.This discovery will aid in the improvement of a potential therapy for muscular dystrophy based on the systemic delivery of MDSCs.

View Article: PubMed Central - PubMed

Affiliation: Department of Neurological Sciences, Stem Cell Laboratory, University of Milan, Padiglione Ponti, Ospedale Policlinico, via Francesco Sforza 35, 20122 Milan, Italy. torrenteyvan@hotmail.com

ABSTRACT
Attempts to repair muscle damage in Duchenne muscular dystrophy (DMD) by transplanting skeletal myoblasts directly into muscles are faced with the problem of the limited migration of these cells in the muscles. The delivery of myogenic stem cells to the sites of muscle lesions via the systemic circulation is a potential alternative approach to treat this disease. Muscle-derived stem cells (MDSCs) were obtained by a MACS(R) multisort method. Clones of MDSCs, which were Sca-1+/CD34-/L-selectin+, were found to adhere firmly to the endothelium of mdx dystrophic muscles after i.v. or i.m. injections. The subpopulation of Sca-1+/CD34- MDSCs expressing L-selectin was called homing MDSCs (HMDSCs). Treatment of HMDSCs with antibodies against L-selectin prevented adhesion to the muscle endothelium. Importantly, we found that vascular endothelium from striate muscle of young mdx mice expresses mucosal addressin cell adhesion molecule-1 (MAdCAM-1), a ligand for L-selectin. Our results showed for the first time that the expression of the adhesion molecule L-selectin is important for muscle homing of MDSCs. This discovery will aid in the improvement of a potential therapy for muscular dystrophy based on the systemic delivery of MDSCs.

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We tested the adhesion of the Sca-1+/CD34−/L-selectin+ positive cells (HMDSCs) to muscle blood vessels by intravital microscopy. Boluses of 5 × 105 fluorescently (BCECF) labeled cells were injected into the quadriceps muscle, and their adhesion to the pectoral muscle vessels was recorded. To improve contrast between the intra- and extravascular compartments, the animals were injected intravenously with a low dose of FITC–dextran. The time course study, i.e., before (a), during (b), and after (c and d) i.m. injections, indicates a significant migration of the HMDSCs within the injected muscle. β-Gal staining of ROSA26-derived HMDSCs showed the presence of injected cells along the injection site (e), near myofibers, bleeding vessels (f), and arterioles (g). Moreover, after i.m. injections in quadriceps, the HMDSCs were able to migrate to the blood stream and then adhered to the endothelial lining of several distant muscles, such as pectoralis (h–m). Arrows in i, k, and m indicate migrated BCECF-labeled cells (bright dots) in the perivascular space of a muscle venule 1 h after injection.
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fig2: We tested the adhesion of the Sca-1+/CD34−/L-selectin+ positive cells (HMDSCs) to muscle blood vessels by intravital microscopy. Boluses of 5 × 105 fluorescently (BCECF) labeled cells were injected into the quadriceps muscle, and their adhesion to the pectoral muscle vessels was recorded. To improve contrast between the intra- and extravascular compartments, the animals were injected intravenously with a low dose of FITC–dextran. The time course study, i.e., before (a), during (b), and after (c and d) i.m. injections, indicates a significant migration of the HMDSCs within the injected muscle. β-Gal staining of ROSA26-derived HMDSCs showed the presence of injected cells along the injection site (e), near myofibers, bleeding vessels (f), and arterioles (g). Moreover, after i.m. injections in quadriceps, the HMDSCs were able to migrate to the blood stream and then adhered to the endothelial lining of several distant muscles, such as pectoralis (h–m). Arrows in i, k, and m indicate migrated BCECF-labeled cells (bright dots) in the perivascular space of a muscle venule 1 h after injection.

Mentions: To identify which cells within the MDSCs were capable of muscle homing, the MDSCs were separated into subpopulations (i.e., Sca-1−/CD34+, Sca-1+/CD34+, and Sca-1+/CD34−) by immunomagnetic selection (MACS® multisort). We then tested the adhesion to muscle blood vessels of these three subpopulations of MDSCs. Boluses of 5 × 105 fluorescently labeled cells were injected into the tail vein or into the quadriceps muscle, and their adhesion to the pectoral muscle vessels was recorded. After i.v. or i.m. injections, no vascular interaction was observed for the Sca-1−CD34+ subpopulation. A few Sca-1+/CD34+ cells interacted with muscle capillaries after i.v. injection but not after i.m. injection. The Sca-1+/CD34− cells were clearly distinguishable from the other two MDSC subpopulations by their significant migration within injected quadriceps after single injection (Fig. 2, c and d). The β-gal staining of muscles injected with Sca-1+/CD34− MDSCs derived from ROSA26 mice confirmed the presence of donor cells spread within myofibers and near muscle capillaries and arterioles (Fig. 2, e–g). Moreover, after i.m. injections, these cells were able to migrate from the injected muscle tissue to the blood stream and then adhered to the endothelial lining of several distant muscles (Fig. 2, h–m).


Identification of a putative pathway for the muscle homing of stem cells in a muscular dystrophy model.

Torrente Y, Camirand G, Pisati F, Belicchi M, Rossi B, Colombo F, El Fahime M, Caron NJ, Issekutz AC, Constantin G, Tremblay JP, Bresolin N - J. Cell Biol. (2003)

We tested the adhesion of the Sca-1+/CD34−/L-selectin+ positive cells (HMDSCs) to muscle blood vessels by intravital microscopy. Boluses of 5 × 105 fluorescently (BCECF) labeled cells were injected into the quadriceps muscle, and their adhesion to the pectoral muscle vessels was recorded. To improve contrast between the intra- and extravascular compartments, the animals were injected intravenously with a low dose of FITC–dextran. The time course study, i.e., before (a), during (b), and after (c and d) i.m. injections, indicates a significant migration of the HMDSCs within the injected muscle. β-Gal staining of ROSA26-derived HMDSCs showed the presence of injected cells along the injection site (e), near myofibers, bleeding vessels (f), and arterioles (g). Moreover, after i.m. injections in quadriceps, the HMDSCs were able to migrate to the blood stream and then adhered to the endothelial lining of several distant muscles, such as pectoralis (h–m). Arrows in i, k, and m indicate migrated BCECF-labeled cells (bright dots) in the perivascular space of a muscle venule 1 h after injection.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC2172686&req=5

fig2: We tested the adhesion of the Sca-1+/CD34−/L-selectin+ positive cells (HMDSCs) to muscle blood vessels by intravital microscopy. Boluses of 5 × 105 fluorescently (BCECF) labeled cells were injected into the quadriceps muscle, and their adhesion to the pectoral muscle vessels was recorded. To improve contrast between the intra- and extravascular compartments, the animals were injected intravenously with a low dose of FITC–dextran. The time course study, i.e., before (a), during (b), and after (c and d) i.m. injections, indicates a significant migration of the HMDSCs within the injected muscle. β-Gal staining of ROSA26-derived HMDSCs showed the presence of injected cells along the injection site (e), near myofibers, bleeding vessels (f), and arterioles (g). Moreover, after i.m. injections in quadriceps, the HMDSCs were able to migrate to the blood stream and then adhered to the endothelial lining of several distant muscles, such as pectoralis (h–m). Arrows in i, k, and m indicate migrated BCECF-labeled cells (bright dots) in the perivascular space of a muscle venule 1 h after injection.
Mentions: To identify which cells within the MDSCs were capable of muscle homing, the MDSCs were separated into subpopulations (i.e., Sca-1−/CD34+, Sca-1+/CD34+, and Sca-1+/CD34−) by immunomagnetic selection (MACS® multisort). We then tested the adhesion to muscle blood vessels of these three subpopulations of MDSCs. Boluses of 5 × 105 fluorescently labeled cells were injected into the tail vein or into the quadriceps muscle, and their adhesion to the pectoral muscle vessels was recorded. After i.v. or i.m. injections, no vascular interaction was observed for the Sca-1−CD34+ subpopulation. A few Sca-1+/CD34+ cells interacted with muscle capillaries after i.v. injection but not after i.m. injection. The Sca-1+/CD34− cells were clearly distinguishable from the other two MDSC subpopulations by their significant migration within injected quadriceps after single injection (Fig. 2, c and d). The β-gal staining of muscles injected with Sca-1+/CD34− MDSCs derived from ROSA26 mice confirmed the presence of donor cells spread within myofibers and near muscle capillaries and arterioles (Fig. 2, e–g). Moreover, after i.m. injections, these cells were able to migrate from the injected muscle tissue to the blood stream and then adhered to the endothelial lining of several distant muscles (Fig. 2, h–m).

Bottom Line: The subpopulation of Sca-1+/CD34- MDSCs expressing L-selectin was called homing MDSCs (HMDSCs).Importantly, we found that vascular endothelium from striate muscle of young mdx mice expresses mucosal addressin cell adhesion molecule-1 (MAdCAM-1), a ligand for L-selectin.This discovery will aid in the improvement of a potential therapy for muscular dystrophy based on the systemic delivery of MDSCs.

View Article: PubMed Central - PubMed

Affiliation: Department of Neurological Sciences, Stem Cell Laboratory, University of Milan, Padiglione Ponti, Ospedale Policlinico, via Francesco Sforza 35, 20122 Milan, Italy. torrenteyvan@hotmail.com

ABSTRACT
Attempts to repair muscle damage in Duchenne muscular dystrophy (DMD) by transplanting skeletal myoblasts directly into muscles are faced with the problem of the limited migration of these cells in the muscles. The delivery of myogenic stem cells to the sites of muscle lesions via the systemic circulation is a potential alternative approach to treat this disease. Muscle-derived stem cells (MDSCs) were obtained by a MACS(R) multisort method. Clones of MDSCs, which were Sca-1+/CD34-/L-selectin+, were found to adhere firmly to the endothelium of mdx dystrophic muscles after i.v. or i.m. injections. The subpopulation of Sca-1+/CD34- MDSCs expressing L-selectin was called homing MDSCs (HMDSCs). Treatment of HMDSCs with antibodies against L-selectin prevented adhesion to the muscle endothelium. Importantly, we found that vascular endothelium from striate muscle of young mdx mice expresses mucosal addressin cell adhesion molecule-1 (MAdCAM-1), a ligand for L-selectin. Our results showed for the first time that the expression of the adhesion molecule L-selectin is important for muscle homing of MDSCs. This discovery will aid in the improvement of a potential therapy for muscular dystrophy based on the systemic delivery of MDSCs.

Show MeSH
Related in: MedlinePlus