Establishment of human Ig producing heterohybridomas by fusion of mouse myeloma cells with human lymphocytes derived from peripheral blood, bone marrow, spleen, lymph node, and synovial fluid. Effect of polyclonal prestimulation and cryopreservation.
50 fusion experiments were carried out to analyse heterohybridization efficiencies on mouse myeloma cells of the P3 X63 Ag8/653 line with human lymphocytes derived from peripheral blood, bone marrow, lymph node, spleen or synovial fluid. We found higher yields of growing and human Ig-producing hybridoma lines when lymphocytes from spleen or lymph node were fused. Although primary hybridomas could be established from fusions with bone marrow-derived cells, only in nine out of 1616 initially seeded wells was Ig production registered. Four fusions using immune cells from synovial fluid were made without success. Independently of the source of lymphocytes pokeweed mitogen (PWM) prestimulation had no enhancing effect on the percentage of wells with cell growth and this did not alter the IgM:IgG ratio in primary hybridomas (9:1), although cells from all compartments used here (with the exception of bone marrow cells) could be stimulated with PWM to produce both IgG and IgM in cultures. Cryopreserved lymphocytes from different sources could be used for fusions with comparable results registered for the fresh material.
Jahn
S
S
Institute for Medical Immunology, Humboldt-University, Berlin, G.D.R.
Grunow
R
R
Kiessig
S T
ST
Specht
U
U
Matthes
H
H
Hiepe
F
F
Hlinak
A
A
Von Baehr
R
R
D000818:Animals; D001854:Bone Marrow Cells; D002459:Cell Fusion; D005615:Freezing; D006801:Humans; D006825:Hybridomas / Q000166:cytology / Q000276:immunology*; D007074:Immunoglobulin G / Q000096:biosynthesis; D007075:Immunoglobulin M / Q000096:biosynthesis; D008198:Lymph Nodes / Q000166:cytology; D008213:Lymphocyte Activation; D008214:Lymphocytes / Q000166:cytology / Q000276:immunology*; D051379:Mice; D011043:Pokeweed Mitogens / Q000494:pharmacology; D013154:Spleen / Q000166:cytology; D013582:Synovial Fluid / Q000166:cytology
D007074:Immunoglobulin G; D007075:Immunoglobulin M; D011043:Pokeweed Mitogens