Purification of human primary neuroblastomas by magnetic beads and their in vitro culture

  • Authors:
    • Hiroko Nakanishi
    • Toshinori Ozaki
    • Youko Nakamura
    • Kohei Hashizume
    • Tadashi Iwanaka
    • Akira Nakagawara
  • View Affiliations

  • Published online on: June 1, 2007     https://doi.org/10.3892/or.17.6.1315
  • Pages: 1315-1320
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Abstract

In the present study we have successfully isolated neuroblastoma cells from primary human neuroblastoma tissues by using a magnetic bead-mediated purification system. Since primary neuroblastoma tissues contained CD3- and CD19-positive lymphocytes, total cell suspensions were prepared and incubated with magnetic beads coated with anti-CD3 or with anti-CD19 antibody. After magnetic separation, unbound materials were recovered and analyzed by immunohistochemical staining for NB84, one of the neuroblastoma markers. Immunohistochemical and FACS analyses demonstrated that NB84-positive cells were enriched in the unbound fraction. Subsequently, unbound materials were seeded on cell culture plates and maintained at 37°C overnight. After incubation, non-adherent cells were collected and stained with anti-NB84 antibody. Under our experimental conditions, a significant increase in the number of NB84-positive cells was observed. Furthermore, our purified NB84-positive cells responded to all-trans retinoic acid and nerve growth factor better than the initial primary cells. Collectively, our present results suggest that magnetic bead-mediated purification enriches neuroblastoma cells which retain their biological properties.

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June 2007
Volume 17 Issue 6

Print ISSN: 1021-335X
Online ISSN:1791-2431

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Spandidos Publications style
Nakanishi H, Ozaki T, Nakamura Y, Hashizume K, Iwanaka T and Nakagawara A: Purification of human primary neuroblastomas by magnetic beads and their in vitro culture. Oncol Rep 17: 1315-1320, 2007.
APA
Nakanishi, H., Ozaki, T., Nakamura, Y., Hashizume, K., Iwanaka, T., & Nakagawara, A. (2007). Purification of human primary neuroblastomas by magnetic beads and their in vitro culture. Oncology Reports, 17, 1315-1320. https://doi.org/10.3892/or.17.6.1315
MLA
Nakanishi, H., Ozaki, T., Nakamura, Y., Hashizume, K., Iwanaka, T., Nakagawara, A."Purification of human primary neuroblastomas by magnetic beads and their in vitro culture". Oncology Reports 17.6 (2007): 1315-1320.
Chicago
Nakanishi, H., Ozaki, T., Nakamura, Y., Hashizume, K., Iwanaka, T., Nakagawara, A."Purification of human primary neuroblastomas by magnetic beads and their in vitro culture". Oncology Reports 17, no. 6 (2007): 1315-1320. https://doi.org/10.3892/or.17.6.1315