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

Oncol Rep. 2007 Jun;17(6):1315-20.

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 degrees 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.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Antigens, CD19 / analysis
  • CD3 Complex / analysis
  • Cell Culture Techniques / methods*
  • Cell Separation / methods*
  • Child
  • Child, Preschool
  • Female
  • Ferrosoferric Oxide / chemistry
  • Flow Cytometry
  • Humans
  • Immunohistochemistry
  • Infant
  • Lymphocytes / chemistry
  • Male
  • Microspheres*
  • Neuroblastoma / pathology*
  • Peripheral Nervous System Neoplasms / pathology*
  • Tumor Cells, Cultured

Substances

  • Antigens, CD19
  • CD3 Complex
  • Ferrosoferric Oxide