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Engraftment of human mesenchymal stem cells in a rat photothrombotic cerebral infarction model : comparison of intra-arterial and intravenous infusion using MRI and histological analysis.

Byun JS, Kwak BK, Kim JK, Jung J, Ha BC, Park S - J Korean Neurosurg Soc (2013)

Bottom Line: In IA group, dark signals in peri-lesional zone were more prominent compared with IV group.SWI showed largest dark signal followed by T2(*)WI and T2WI in both IA and IV groups.In a rat photothrombotic model of ischemic stroke, selective IA administration of human mesenchymal stem cells is more effective than IV administration.

View Article: PubMed Central - PubMed

Affiliation: Department of Radiology, Chung-Ang University College of Medicine, Seoul, Korea.

ABSTRACT

Objective: This study aimed to evaluate the hypotheses that administration routes [intra-arterial (IA) vs. intravenous (IV)] affect the early stage migration of transplanted human bone marrow-derived mesenchymal stem cells (hBM-MSCs) in acute brain infarction.

Methods: Male Sprague-Dawley rats (n=40) were subjected to photothrombotic infarction. Three days after photothrombotic infarction, rats were randomly allocated to one of four experimental groups [IA group : n=12, IV group : n=12, superparamagnetic iron oxide (SPIO) group : n=8, control group : n=8]. All groups were subdivided into 1, 6, 24, and 48 hours groups according to time point of sacrifice. Magnetic resonance imaging (MRI) consisting of T2 weighted image (T2WI), T2(*) weighted image (T2(*)WI), susceptibility weighted image (SWI), and diffusion weighted image of rat brain were obtained prior to and at 1, 6, 24, and 48 hours post-implantation. After final MRI, rats were sacrificed and grafted cells were analyzed in brain and lung specimen using Prussian blue and immunohistochemical staining.

Results: Grafted cells appeared as dark signal intensity regions at the peri-lesional zone. In IA group, dark signals in peri-lesional zone were more prominent compared with IV group. SWI showed largest dark signal followed by T2(*)WI and T2WI in both IA and IV groups. On Prussian blue staining, IA administration showed substantially increased migration and a large number of transplanted hBM-MSCs in the target brain than IV administration. The Prussian blue-positive cells were not detected in SPIO and control groups.

Conclusion: In a rat photothrombotic model of ischemic stroke, selective IA administration of human mesenchymal stem cells is more effective than IV administration. MRI and histological analyses revealed the time course of cell migration, and the numbers and distribution of hBM-MSCs delivered into the brain.

No MeSH data available.


Related in: MedlinePlus

In vitro cell staining. A : Non-labeled human mesenchymal stem cell, Prussian blue staining, original magnification ×400. B : Labeled hMSC, Prussian blue staining, original magnification ×400. C : Combined staining with antimitochondrial antibody-Prussian, original magnification ×400.
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Figure 1: In vitro cell staining. A : Non-labeled human mesenchymal stem cell, Prussian blue staining, original magnification ×400. B : Labeled hMSC, Prussian blue staining, original magnification ×400. C : Combined staining with antimitochondrial antibody-Prussian, original magnification ×400.

Mentions: We used a clinically approved SPIO (Feridex IV, ferumoxide; Advanced Susceptibility Inc., Cambridge, MA, USA). The SPIO are characterized by a 5-nm iron core and hydrodynamic diameter of 80-90 nm, including the dextran coating51). Ferumoxide is provided as an SPIO solution with a total iron content of 11.2 mg/mL and is approved for clinical use in liver imaging. By reducing the transverse relaxation time (T2) on T2-weighted images, the SPIO nanoparticles cause labeled cells to appear as regions of dark signal intensity. To facilitate endocytosis of SPIO by the hBM-MSCs, the polyamine poly-L-lysine hydrobromide (PLL; Sigma-Aldrich Corp., St. Louis, MO, USA) was used37). To obtain a final PLL concentration of 375 ng/mL and 25 ug/mL of SPIO in the medium, the PLL at 750 ng/mL and SPIO at 50 ug/mL were mixed with culture medium and gently shaken for 60 minutes at room temperature. The hBM-MSCs were labeled with SPIO by incubation overnight in medium containing SPIO-PLL complex at 37℃ in a 5% CO2 humidified atmosphere. The viability of the SPIO-labeled hBM-MSCs before transplantation was assessed at greater than 85% by trypan blue exclusion. Before injection into rats, the labeled cells were prepared in phosphate-buffered saline (PBS). Nearly 100% labeling of hBM-MSCs with SPIO was achieved, as shown by Prussian blue and immunohistochemical staining, and cellular MRI performed in vitro (Fig. 1A, B). Mitochondria and iron were detected in the hBM-MSC by combined staining with anti-mitochondrial antibody and Prussian blue (Fig. 1C).


Engraftment of human mesenchymal stem cells in a rat photothrombotic cerebral infarction model : comparison of intra-arterial and intravenous infusion using MRI and histological analysis.

Byun JS, Kwak BK, Kim JK, Jung J, Ha BC, Park S - J Korean Neurosurg Soc (2013)

In vitro cell staining. A : Non-labeled human mesenchymal stem cell, Prussian blue staining, original magnification ×400. B : Labeled hMSC, Prussian blue staining, original magnification ×400. C : Combined staining with antimitochondrial antibody-Prussian, original magnification ×400.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC3921273&req=5

Figure 1: In vitro cell staining. A : Non-labeled human mesenchymal stem cell, Prussian blue staining, original magnification ×400. B : Labeled hMSC, Prussian blue staining, original magnification ×400. C : Combined staining with antimitochondrial antibody-Prussian, original magnification ×400.
Mentions: We used a clinically approved SPIO (Feridex IV, ferumoxide; Advanced Susceptibility Inc., Cambridge, MA, USA). The SPIO are characterized by a 5-nm iron core and hydrodynamic diameter of 80-90 nm, including the dextran coating51). Ferumoxide is provided as an SPIO solution with a total iron content of 11.2 mg/mL and is approved for clinical use in liver imaging. By reducing the transverse relaxation time (T2) on T2-weighted images, the SPIO nanoparticles cause labeled cells to appear as regions of dark signal intensity. To facilitate endocytosis of SPIO by the hBM-MSCs, the polyamine poly-L-lysine hydrobromide (PLL; Sigma-Aldrich Corp., St. Louis, MO, USA) was used37). To obtain a final PLL concentration of 375 ng/mL and 25 ug/mL of SPIO in the medium, the PLL at 750 ng/mL and SPIO at 50 ug/mL were mixed with culture medium and gently shaken for 60 minutes at room temperature. The hBM-MSCs were labeled with SPIO by incubation overnight in medium containing SPIO-PLL complex at 37℃ in a 5% CO2 humidified atmosphere. The viability of the SPIO-labeled hBM-MSCs before transplantation was assessed at greater than 85% by trypan blue exclusion. Before injection into rats, the labeled cells were prepared in phosphate-buffered saline (PBS). Nearly 100% labeling of hBM-MSCs with SPIO was achieved, as shown by Prussian blue and immunohistochemical staining, and cellular MRI performed in vitro (Fig. 1A, B). Mitochondria and iron were detected in the hBM-MSC by combined staining with anti-mitochondrial antibody and Prussian blue (Fig. 1C).

Bottom Line: In IA group, dark signals in peri-lesional zone were more prominent compared with IV group.SWI showed largest dark signal followed by T2(*)WI and T2WI in both IA and IV groups.In a rat photothrombotic model of ischemic stroke, selective IA administration of human mesenchymal stem cells is more effective than IV administration.

View Article: PubMed Central - PubMed

Affiliation: Department of Radiology, Chung-Ang University College of Medicine, Seoul, Korea.

ABSTRACT

Objective: This study aimed to evaluate the hypotheses that administration routes [intra-arterial (IA) vs. intravenous (IV)] affect the early stage migration of transplanted human bone marrow-derived mesenchymal stem cells (hBM-MSCs) in acute brain infarction.

Methods: Male Sprague-Dawley rats (n=40) were subjected to photothrombotic infarction. Three days after photothrombotic infarction, rats were randomly allocated to one of four experimental groups [IA group : n=12, IV group : n=12, superparamagnetic iron oxide (SPIO) group : n=8, control group : n=8]. All groups were subdivided into 1, 6, 24, and 48 hours groups according to time point of sacrifice. Magnetic resonance imaging (MRI) consisting of T2 weighted image (T2WI), T2(*) weighted image (T2(*)WI), susceptibility weighted image (SWI), and diffusion weighted image of rat brain were obtained prior to and at 1, 6, 24, and 48 hours post-implantation. After final MRI, rats were sacrificed and grafted cells were analyzed in brain and lung specimen using Prussian blue and immunohistochemical staining.

Results: Grafted cells appeared as dark signal intensity regions at the peri-lesional zone. In IA group, dark signals in peri-lesional zone were more prominent compared with IV group. SWI showed largest dark signal followed by T2(*)WI and T2WI in both IA and IV groups. On Prussian blue staining, IA administration showed substantially increased migration and a large number of transplanted hBM-MSCs in the target brain than IV administration. The Prussian blue-positive cells were not detected in SPIO and control groups.

Conclusion: In a rat photothrombotic model of ischemic stroke, selective IA administration of human mesenchymal stem cells is more effective than IV administration. MRI and histological analyses revealed the time course of cell migration, and the numbers and distribution of hBM-MSCs delivered into the brain.

No MeSH data available.


Related in: MedlinePlus