J Korean Neurosurg Soc.  2010 Nov;48(5):391-398. 10.3340/jkns.2010.48.5.391.

In vivo Tracking of Transplanted Bone Marrow-Derived Mesenchymal Stem Cells in a Murine Model of Stroke by Bioluminescence Imaging

Affiliations
  • 1Department of Neurosurgery, Institute of Catholic Integrative Medicine (ICIM) of Incheon St. Mary's Hospital, The Catholic University of Korea, Incheon, Korea.
  • 2Department of Neurosurgery, Seoul St. Mary's Hospital, The Catholic University of Korea, Seoul, Korea. nslk@catholic.ac.kr
  • 3Department of Neurosurgery, St. Vincent's Hospital, The Catholic University of Korea, Suwon, Korea.

Abstract


OBJECTIVE
This study was designed to validate the cell trafficking efficiency of the in vivo bioluminescence image (BLI) study in the setting of transplantation of the luciferase expressing bone marrow-derived mesenchymal stem cells (BMSC), which were delivered at each different time after transient middle cerebral artery occlusion (MCAO) in a mouse model.
METHODS
Transplanting donor BMSC were prepared by primary cell culture from transgenic mouse expressing luciferase (LUC). Transient focal infarcts were induced in 4-6-week-old male nude mice. The experiment mice were divided into five groups by the time of MSC transplantation : 1) sham-operation group, 2) 2-h group, 3) 1-day group, 4) 3-day group, and 5) 1- week group. BLI for detection of spatial distribution of transplanted MSC was performed by detecting emitted photons. Migration of the transplanted cells to the infarcted area was confirmed by histological examinations. Differences between groups were evaluated by paired t-test.
RESULTS
A focal spot of bioluminescence was observed at the injection site on the next day after transplantation by signal intensity of bioluminescence. After 4 weeks, the mean signal intensities of 2-h, 1-day, 3-day, and 1-week group were 2.6x10(7) +/- 7.4x10(6), 6.1x10(6) +/- 1.2x10(6), 1.7x10(6) +/- 4.4x10(5), and 8.9x10(6) +/- 9.5x10(5), respectively. The 2-h group showed significantly higher signal intensity (p < 0.01). The engrafted BMSC showed around the infarct border zones on immunohistochemical examination. The counts of LUC-positive cells revealed the highest number in the 2-h group, in agreement with the results of BLI experiments (p < 0.01).
CONCLUSION
In this study, the results suggested that the transplanted BMSC migrated to the infarct border zone in BLI study and the higher signal intensity of LUC-positive cells seen in 2 hrs after MSC transplantation in MCAO mouse model. In addition, noninvasive imaging in real time is an ideal method for tracking stem cell transplantation. This method can be widely applied to various research fields of cell transplantation therapy.

Keyword

Stroke; Bone marrow-derived mesenchymal stem cells; Bioluminescence image

MeSH Terms

Animals
Cell Transplantation
Enzyme Multiplied Immunoassay Technique
Humans
Infarction, Middle Cerebral Artery
Luciferases
Male
Mesenchymal Stromal Cells
Mice
Mice, Nude
Mice, Transgenic
Photons
Primary Cell Culture
Stem Cell Transplantation
Stroke
Tissue Donors
Track and Field
Transplants
Luciferases
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