Long-term culture of primary B cells and in vitro expression of an exogenous gene

Immunol Lett. 1995 Sep;47(3):193-7. doi: 10.1016/0165-2478(95)00092-7.

Abstract

Efficient introduction and expression of exogenous genes into primary B cells is very important to study B-cell biology and is essential for gene therapy. These efforts have often been impeded by the lack of availability of a simple culture condition for growth and proliferation of primary B cells as well as the lack of vehicles for efficient introduction of genes of interest. In this communication, we have developed a culture condition that supports the growth of primary B cells from beta-gal-immunized mouse spleen for 30 days without the aid of feeder layers. During this period, B cells secreted polyclonal antibodies into the medium. To study expression of an exogenous reporter gene, human growth hormone (hGH) was introduced into cultured B cells using retroviral vectors. hGH was expressed up to 21 days in the absence of drug selection and the infected cells continued to secrete immunoglobulins into the medium.

MeSH terms

  • Animals
  • B-Lymphocytes / immunology
  • B-Lymphocytes / physiology*
  • Cell Culture Techniques / methods*
  • Gene Expression
  • Genetic Vectors
  • Growth Hormone / genetics*
  • Immunoglobulins / biosynthesis
  • Mice
  • Mice, Inbred BALB C
  • Spleen / cytology*
  • Spleen / immunology
  • Time Factors
  • Transduction, Genetic*

Substances

  • Immunoglobulins
  • Growth Hormone