Establishment of a Multipotent Neural Stem Cell Line from the Adult Mouse Cerebrum / 대한척추외과학회지

ABSTRACT

STUDY DESIGN: Establishment of a multipotent neural stem cell line from the adult mouse cerebrum. OBJECTIVES: To establish a daughter cell line, B2A1, from B2 cells through the limiting dilution method, and to determine if the cells have the characteristics of neural stem cell (NSCs) using immunocytochemistry and RT-PCR methods. SUMMARY AND LITERATURE REVIEW In the development of NSCs, differentiated organ or tissue-derived multipotent stem cells have attracted considerable interest because of the lack of ethical issues. Previously, a glial precursor cell line (B2 cells) was generated from the primary cultures of oligodendrocytes/ astrocytes in an adult BALB/c mouse brain. These cells exhibited the cell-type specific markers for immature neuroectodermal cells, astrocytes, and oligodendrocytes in serum-contained media. MATERIALS AND METHODS: The primary cultures of oligodendrocytes/astrocytes were established from the whole brains of 12 to 16-week-old BALB/c mice from either gender. After 6 months with 25 serial passages, the culture consisted of a morphologically homogeneous cell population, which was designated as B2 cells. A subclone, B2A1, was isolated from B2 cells through two consecutive limiting dilutions. RESULTS: More than 90% of B2A1 cells showed immunopositivity for nestin, a specific marker for NSC. The cells also showed immunopositivity for the neuronal, astrocytic and oligodendroglial markers. These cells expressed the genotypic mRNA messages for both neural progenitor cells and differentiated neuronoglial cells. These positive immunocytochemical reactions and mRNA messages for neuronoglial cells varied according to the extrinsic growth factors used. However, the treatment of extrinsic growth factors did not produce any significant differences in the nestin-immunopositive cells. CONCLUSIONS: B2A1 cells have the immunocytochemical and cytogenetic properties of NSCs, and the capacity to differentiate into neuronoglial cells.