Generation of chromosome-specific monoclonal antibodies using in vitro differentiated trans-chromosomic (TC) mouse ES cells

¹ Pharmaceutical Research Laboratories, Pharmaceutical Division, KIRIN Brewery Co., Ltd. Takasaki, Gunma, Japan, and Department of Biomedical Science, Institute of Regenerative Medicine and Biofunction, Graduate School of Medicine, Tottori University, Yonago, Tottori, Japan
² Pharmaceutical Research Laboratories, Pharmaceutical Division, KIRIN Brewery Co., Ltd. Takasaki, Gunma, Japan
³ Department of Biomedical Science, Institute of Regenerative Medicine and Biofunction, Graduate School of Medicine, Tottori University, Yonago, Tottori, Japan

^* To whom correspondence should be addressed. E-mail: ktomizuka{at}kirin.co.jp.

	Abstract

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 ES cells. In this report, we examined the use of in vitro differentiated trans-chromosomic mouse ES (TC-ES) cells 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-ES cells resulted in two chromosome-specific MoAbs, h4-neural1 and h11-neural1, respectively. The staining profiles of differentiated TC-ES cells and human embryonic carcinoma (EC) cells with these MoAbs were similar to the expression profile of nestin, a well-characterized intracellular marker for neural progenitor cells (NPCs). 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 human ES cells.