Fibroblast-dependent growth of mouse mast cells in vitro: Duplication of mast cell depletion in mutant mice of w/wv genotype

✍ Scribed by Jun Fujita; Hiroki Nakayama; Hitoshi Onoue; Yuzuru Kanakura; Toru Nakano; Hidekazu Asai; Shun-Ichi Takeda; Tasuku Honjo; Yukihiko Kitamura

• Publisher: John Wiley and Sons
• Year: 1988
• Tongue: English
• Weight: 891 KB
• Volume: 134
• Category: Article
• ISSN: 0021-9541
• DOI: 10.1002/jcp.1041340109

✦ Synopsis

In spite of the apparent depletion of mast cells in tissues of mutant mice of W/W" genotype, cells with many features of mast cells do develop when bone marrow cells of W/W" 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/WVor congenic + I + mice, and then cocultured 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 coculture 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/W" mice and might be useful for biochemical and molecular analysis of the gene product(s) encoded at the W locus.