Generation and characterization of monoclonal antibodies to the neural crest

The generation of monoclonal antibodies (MAbs) specific for quail neural cr est may provide valuable tools for studying the differentiation of embryoni c precursor cells, Unfortunately, relatively few antibodies are available b ecause of the difficulty in obtaining sufficient cells for in vivo immuniza tion strategies, We have overcome this problem by using intrasplenic immuni zation with formaldehyde-fixed cells harvested from neural crest cultures. In addition, booster injections of cultured whole-embryo cells were adminis tered intraperitoneally, Following two fusions, a total of 18 hybridomas we re generated with antibody reactivity to the cytoplasm of neural crest cell s. Furthermore, 32 were reactive against both somite (a noncrest mesodermal control) and crest cultures, whilst 15 were not reactive. Out of those hyb ridomas reactive with neural crest, six designated 160D, 164D, OE, 12E, 120 E and 124E were further characterized. Interestingly MAb supernatants OE, 1 2E, 120E, and 124E exhibited reactivity against some but not all neural cre st cells suggesting that they might recognise subpopulations. Immunoglobuli n isotyping of supernatants revealed that 4 (160D, 164D, OE, and 120E) were IgM and 2 (12E and 124E) were IgG(2b), On assessing their reactivity again st human tissue sections, all six hybridoma supernatants cross-reacted with neuroendocrine cells within appendix, colon and rectum, These MAbs could p rovide novel reagents for the understanding of neural crest development.