Long-term culture of primary B cells and in vitro expression of an exogenous gene.
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.
Kumar
A
A
Molecular Biology Group, Hybritech Incorporated, P.O. Box 269006, San Diego, CA 92196, USA.
Guido
E C
EC
Liu
R S
RS
Saedi
M S
MS
D000818:Animals; D001402:B-Lymphocytes / Q000276:immunology / Q000502:physiology*; D018929:Cell Culture Techniques / Q000379:methods*; D015870:Gene Expression; D005822:Genetic Vectors; D013006:Growth Hormone / Q000235:genetics*; D007136:Immunoglobulins / Q000096:biosynthesis; D051379:Mice; D008807:Mice, Inbred BALB C; D013154:Spleen / Q000166:cytology* / Q000276:immunology; D013997:Time Factors; D014161:Transduction, Genetic
D007136:Immunoglobulins; D013006:Growth Hormone