Open AccessGeneration of Chromosome‐Specific Monoclonal Antibodies Using In Vitro–Differentiated Transchromosomic Mouse Embryonic Stem Cells
Author(s) -
Yanagisawa Ayano,
Endo Chisato,
Okawa Katsuya,
Shitara Shingo,
Kugoh Hiroyuki,
Kakitani Makoto,
Oshimura Mitsuo,
Tomizuka Kazuma
Publication year - 2005
Publication title -
stem cells
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 2.159
H-Index - 229
eISSN - 1549-4918
pISSN - 1066-5099
DOI - 10.1634/stemcells.2004-0369
Subject(s) - biology , embryonic stem cell , microbiology and biotechnology , monoclonal antibody , antigen , stem cell , cellular differentiation , progenitor cell , population , antibody , immunology , genetics , gene , demography , sociology
Monoclonal antibodies (MoAbs) recognizing lineage‐ and stage‐specific human cell‐surface antigens are valuable reagents for the characterization and isolation of various specialized cell populations derived from human embryonic stem cells (hESCs). In this report, we examined the use of in vitro differentiated transchromosomic mouse embryonic stem cells (TC‐ESCs) as immunogens to obtain MoAbs against human cell‐surface antigens. Immunization of a neural‐cell population derived from differentiating human chromosome 4 and 11 TC‐ESCs resulted in two chromosome‐specific MoAbs, h4‐neural1 and h11‐neural1, respectively. The staining profiles of differentiated TC‐ESCs and human embryonic carcinoma cells with these MoAbs were similar to the expression profile of nestin, a well‐characterized intracellular marker for neural progenitor cells. We also described the successful purification and identification of the gene for h4‐neural1 antigen (CD133, 4p15.32) with immunoaffinity chromatography. This procedure may have significant utility in generating MoAbs useful for understanding the mechanism that regulates the in vitro differentiation of hESCs.