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Summary

Abstract

Introduction

Protocol

Representative Results

Discussion

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References

Biology

Intraspinal Cavity Injection of Human Mesenchymal Stem Cells and Tracking their Migration into the Rat Brain

Published: February 3rd, 2021

DOI:

10.3791/62120

1Stem Cell & Regenerative Medicine Institute, Samsung Medical Center, 2Stem Cell Institute, ENCell Co. Ltd, 3Department of Neurosurgery, Samsung Medical Center, Sungkyunkwan University School of Medicine, 4Animal Research and Molecular Imaging Center, Samsung Biomedical Research Institute, Samsung Medical Center, 5Department of Neurology, Samsung Medical Center, Sungkyunkwan University School of Medicine, 6Neuroscience Center, Samsung Medical Center, 7Samsung Alzheimer Research Center, Samsung Medical Center

Several routes of administration can be used to deliver mesenchymal stem cells (MSCs) to the brain. In the present study, MSCs were delivered throughout the neuraxis and brain via intra-spinal cavity injection. MSCs were injected into the spinal cavities of rats, and stem cell migration was tracked and quantified.

Mesenchymal stem cells (MSCs) have been studied for the treatment of various diseases. In neurodegenerative diseases involving defects in both the brain and the spinal cord, the route of administration is very important, because MSCs must migrate to both the brain and the spinal cord. This paper describes a method for administering MSCs into the spinal canal (intraspinal cavity injection) that can target the brain and spinal cord in a rat model. One million MSCs were injected into the spinal canals of rats at the level of lumbar vertebrae 2-3. After administration, the rats were euthanized at 0, 6, and 12 h post-injection. Optical imaging and quantitative real-time polymerase chain reaction (qPCR) were used to track the injected MSCs. The results of the present study demonstrated that MSCs administered via the spinal cavity could be detected subsequently in both the brain and spinal cord at 12 h. Intraspinal cavity injection has the advantage of not requiring general anesthesia and has few side effects. However, the drawback of the low migration rate of MSCs to the brain must be overcome.

Mesenchymal stem cells
Under disease conditions, MSCs secrete disease-specific therapeutic substances via paracrine actions1 that have been reported to regulate immune responses, restore damaged tissues, and remove toxic substances2. Therefore, MSC therapy is considered more effective than single-target therapy in treating multifactorial diseases such as Alzheimer's disease and sarcopenia3,4,5,6. Additionally, in contrast to pharmaceuticals, MSCs have a homing effect,....

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NOTE: This study was approved by the Institutional Animal Care and Use Committee (Approval number: 20170125001, Date: January 25, 2017) of the Samsung Biomedical Research Institute (SBRI) at Samsung Medical Center. As an accredited facility of the Association for Assessment and Accreditation of Laboratory Animal Care International, the SBRI acts in accordance with the guidelines set forth by the Institute of Laboratory Animal Resources.

1. Preparation of human Wharton's jelly-derived MSCs

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To evaluate the efficacy of intraspinal cavity injection of MSCs, DiD-labeled MSCs were used in the present study. Before injecting MSCs into the spinal cavity, the labeling efficacy was assessed in vitro using optical imaging and fluorescence microscopy (Figure 1). After staining the MSCs with the DiD labeling reagent using the procedure described in protocol section 3.1, optical images were taken of the culture plates on which DiD-labeled MSCs were seeded (Figure 1A

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The optimal route of administration for treatment with MSCs should be chosen depending on the target disease, the patient's condition, and the type of drug to be delivered. In cell therapies, including MSC therapy, direct injection of stem cells into the brain or intrathecally via the CSF must be considered as the cells cannot pass through the BBB19. Intraspinal cavity injection is relatively non-invasive and does not cause neuronal damage in the brain, unlike intracerebroventricular injection.......

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This study was supported by grants from the Basic Research Program through the National Research Foundation of South Korea (NRF), funded by the Ministry of Education (NRF-2017R1D1A1B03035940), and a grant from the Korea Health Technology R&D Project through the Korea Health Industry Development Institute (KHIDI), funded by the Ministry of Health & Welfare, Republic of Korea (grant numbers: HI14C3484 and HI18C0560). We would like to thank Editage (www.editage.co.kr) for English language editing.

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Name Company Catalog Number Comments
0.25% Trypsin-EDTA Gibco-invitrogen 25200114 Cell culture
Fetal bovine serum biowest S1520 Culture medium supplement
gentamicin Gibco-invitrogen 15710-072 Culture medium supplement
Gentra Puregene Tissue Kit QIAGEN 158689 gDNA isolation
MEM, no glutamine, no phenol red Gibco 51200038 WJ-MSC fomulation for injection
Miminum Essential Medium alpha Gibco-invitrogen 12571063 WJ-MSC culture medium
Power SYBR Green PCR Master Mix Applied Biosystems 4368577 quantitative real time PCR reagent
QuantStudio 6 Flex Real-Time PCR System Thermo fisher 4485694 quantitative real time PCR
trypan blue Gibco 15250061 Injection
Vybrant DiD Cell-Labeling Solution invitrogen V22887 Stem cell labeling solution
Xenogen IVIS Spectrum system Perkin Elmer 124262 Optical imaging device

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