Generation of chromosome-specific monoclonal antibodies using in vitro-differentiated transchromosomic mouse embryonic stem cells.
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.
Yanagisawa
Ayano
A
Pharmaceutical Research Laboratory, Pharmaceutical Division, Kirin Brewery Co., Ltd., Takasaki-shi, Gunma 370-1295, Japan.
Endo
Chisato
C
Okawa
Katsuya
K
Shitara
Shingo
S
Kugoh
Hiroyuki
H
Kakitani
Makoto
M
Oshimura
Mitsuo
M
Tomizuka
Kazuma
K
D000071916:AC133 Antigen; D000818:Animals; D000911:Antibodies, Monoclonal / Q000096:biosynthesis*; D015703:Antigens, CD / Q000235:genetics / Q000378:metabolism*; D000954:Antigens, Surface / Q000235:genetics / Q000378:metabolism; D019002:CD56 Antigen / Q000235:genetics / Q000378:metabolism*; D002454:Cell Differentiation; D002478:Cells, Cultured; D033481:Chromosomes, Mammalian / Q000235:genetics / Q000276:immunology*; D033041:Embryo Research; D006023:Glycoproteins / Q000235:genetics / Q000378:metabolism*; D006801:Humans; D051379:Mice; D009474:Neurons / Q000166:cytology; D010455:Peptides / Q000235:genetics / Q000378:metabolism*; D013234:Stem Cells / Q000166:cytology*; D019076:Transgenes
D000071916:AC133 Antigen; D000911:Antibodies, Monoclonal; D015703:Antigens, CD; D000954:Antigens, Surface; D019002:CD56 Antigen; D006023:Glycoproteins; C501063:Ncam1 protein, mouse; C000606023:PROM1 protein, human; D010455:Peptides; C000606024:Prom1 protein, mouse