Fibroblast-dependent growth of mouse mast cells in vitro: duplication of mast cell depletion in mutant mice of W/Wv genotype.
In spite of the apparent depletion of mast cells in tissues of mutant mice of W/Wv genotype, cells with many features of mast cells do develop when bone marrow cells of W/Wv mice are cultured in the presence of pokeweed mitogen-stimulated spleen cell-conditioned medium (PWM-SCM). In order to resolve this discrepancy and facilitate the analysis of the W mutation, we attempted to establish an in vitro system in which the in vivo defect of W/Wv mice can be reproduced. Cultured mast cells (CMC) were developed from bone marrow cells of either W/Wv or congenic +/+ mice, and then co-cultured with NIH/3T3 mouse fibroblasts in media supplemented only with fetal calf serum (i.e., in the absence of PWM-SCM). Under this condition, CMC from +/+ mice continued to divide and were maintained for more than 4 weeks. The supportive effect of NIH/3T3 cells required close-range interactions with CMC and was not due to synthesis of the known mast cell growth factors, interleukins 3 and 4. By contrast, CMC from W/Wv mice were not maintained, and the number of mast cells remaining after 4 weeks of co-culture was only 1% of the normal +/+ counterparts. Thus, the humoral factor-independent and cell contact-dependent system presented here revealed the intrinsic defects in growth and differentiation of CMC derived from W/Wv mice and might be useful for biochemical and molecular analysis of the gene product(s) encoded at the W locus.
Fujita
J
J
Institute for Cancer Research, Osaka University Medical School, Japan.
Nakayama
H
H
Onoue
H
H
Kanakura
Y
Y
Nakano
T
T
Asai
H
H
Takeda
S
S
Honjo
T
T
Kitamura
Y
Y
D000818:Animals; D002450:Cell Communication; D002455:Cell Division; D002460:Cell Line; D003584:Cytological Techniques; D005347:Fibroblasts / Q000378:metabolism / Q000502:physiology*; D005838:Genotype; D006493:Heparin / Q000096:biosynthesis; D007377:Interleukin-3 / Q000096:biosynthesis; D015847:Interleukin-4; D007378:Interleukins / Q000096:biosynthesis; D008407:Mast Cells / Q000166:cytology* / Q000378:metabolism; D051379:Mice; D008817:Mice, Mutant Strains / Q000033:anatomy & histology*
D007377:Interleukin-3; D007378:Interleukins; D015847:Interleukin-4; D006493:Heparin