J Korean Neurosurg Soc.  2005 Aug;38(2):121-125.

The Cell Survival and Differentiation after Transplantation, Which Harvest from Adult Rat Brain by High-speed Centrifugation Method

Affiliations
  • 1Department of Neurosurgery, Our Lady Mercy Hospital, College of Medicine The Catholic University of Korea, Incheon, Korea.
  • 2Department of Neurosurgery, Uijongbu St. Mary's Hospital, College of Medicine The Catholic University of Korea, Uijeongbu, Korea. yooman@catholic.ac.kr

Abstract


OBJECTIVE
Many recent reports have shown that the mature mammalian brain harbors multipotent stem cells, rendering the brain capable of generating new neurons and glia throughout life. Harvested stem cells from an adult rat are transplanted in order to evaluate the cell survival and differentiation. METHODS: Using a percoll gradient with a high speed centrifugation method, we isolate neural stem/progenitor cells were isolated from the subventricular zone(SVZ) of a syngeneic adult Fisher 344 rats brain. For 14days expansion, the cultured cells comprised of a heterogeneous population with the majority of cells expressing nestin and/or GFAP. After expanding the SVZ cells in the presence of basic fibroblast growth factor-2, and transplanting then into the hippocampus of normal rats, the survival and differentiation of those cells were examined. For transplantation, the cultured cells were labeled with BrdU two days prior to use. In order to test their survival, the cells were transplanted into the dorsal hippocampus of normal adult Fisher 344 rats. RESULTS: The preliminary data showed that at 7days after transplantation, BrdU+ transplanted cells were observed around the injection deposition sites. Immuno-fluorescent microscopy revealed that the cells co-expressed BrdU+ and neuronal marker beta-tubulin III. CONCLUSION: The data demonstrate that the in vitro expanded SVZ cells can survive in a heterotypic environment and develop a neuronal phenotype in the neurogenic region. However more research will be needed to examine the longer survival time points and quantifying the differentiation in the transplanted cells in an injured brain environment.

Keyword

Neural stem cell; Transplantation; Percoll solution; High-speed centrifugation; Subventricular zone; Hippocampus

MeSH Terms

Adult*
Animals
Brain*
Bromodeoxyuridine
Cell Survival*
Cells, Cultured
Centrifugation*
Fibroblast Growth Factor 2
Hippocampus
Humans
Microscopy
Multipotent Stem Cells
Nestin
Neural Stem Cells
Neuroglia
Neurons
Phenotype
Rats*
Stem Cells
Transplantation
Tubulin
Bromodeoxyuridine
Fibroblast Growth Factor 2
Tubulin
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