Int J Stem Cells.  2015 Nov;8(2):191-199. 10.15283/ijsc.2015.8.2.191.

A Novel Cell Therapy Method for Recovering after Brain Stroke in Rats

• Hosseini S
• Farahmandnia M
• Kazemi S
• Shakibajahromi B
• Sarvestani FS
• Khodabande Z

• KMID: 2380813
• DOI: http://doi.org/10.15283/ijsc.2015.8.2.191

Abstract

BACKGROUND
Nowadays, stroke leads to a significant part of the adult mortality and morbidity and also it could result in some neurological deficits in the patients' lives. Cell therapy has opened a new approach to treat the brain ischemia and reduce its terrible effects on the patients' lives. There are several articles which show that the cell therapy could be beneficial for treating brain stroke. In this study, we have planned to present a new cell therapy method for stroke by administration of Mesenchymal stem cells and differentiated neural stem cells without astrocytes. METHOD AND MATERIALS: The Mesenchymal stem cells were isolated from tibia and femur of a 250~300 g rat and they were cultured in DMEM/F12, 10% fetal bovine serum, 1% Pen/Strep. Neural stem cells were isolated from 14 days rat embryo ganglion eminence and were cultured in NSA media containing Neurobasal, 2% B27, bFGF 10 ng/ml and EGF 20 ng/ml after 5 days they formed some neurospheres. The isolated neural stem cells were differentiated to neural lineages by adding 5% fetal bovine serum to their culture media. After 48 hours the astrocytes were depleted by using MACS kit.
RESULTS
The group that received Mesenchymal stem cells systemically and differentiated neural stem cells without astrocytes had the best neurological outcomes and the least infarct volume and apoptosis. It could be understood that this cell therapy method might cause almost full recovery after brain stoke.
CONCLUSION
Using combination cell therapy with Mesenchymal stem cells and differentiated neural stem cells with removed astrocyte could provide a novel method for curing brain stroke.

Keyword

Neural stem cells; Mesenchymal stem cells; Brain stroke; Astrocyte

MeSH Terms

Adult
Animals
Apoptosis
Astrocytes
Brain Ischemia
Brain*
Cell- and Tissue-Based Therapy*
Culture Media
Embryonic Structures
Epidermal Growth Factor
Femur
Ganglion Cysts
Humans
Mesenchymal Stromal Cells
Mortality
Neural Stem Cells
Rats*
Stroke*
Tibia
Culture Media
Epidermal Growth Factor

Figure

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  Fig. 1 Rat Mesenchymal stem cells.

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  Fig. 2 Rat Mesenchymal stem cell immunocytochemistry for CD44, CD90, CD45 and CD34.

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  Fig. 3 Mesenchymal stem cells differentiation to adipocyte and osteocyte.

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  Fig. 4 Neurospheres 5 days after neural stem cell culture.

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  Fig. 5 Differentiated neural stem cells.

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  Fig. 6 Neural Stem Cell Differentiation and immunocytochemistry with GFAP and β-tubulin III antibody.

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  Fig. 7 Flow cytometry for assessing depletion of astrocytes. The blue line is the GFAP positive cells.

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  Fig. 8 Neurological scores in 28 days. The data was analyzed with One-way ANOVA and the p value is less than 0.05. The data between groups was analyzed with Tukey’s multiple comparison tests.

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  Fig. 9 H&E staining and ischemic area volume. The data was analyzed with One-way ANOVA and the p value is less than 0.05. The data between groups was analyzed with Tukey’s multiple comparison tests.

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  Fig. 10 H&E staining for brain tissue.

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  Fig. 11 Caspase 3 activity. The data was analyzed with One-way ANOVA and the p value is less than 0.05. The data between groups was analyzed with Tukey’s multiple comparison tests.

Reference

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