Selection against undifferentiated human embryonic stem cells by a cytotoxic antibody recognizing podocalyxin-like protein-1.
Future therapeutic applications of differentiated human embryonic stem cells (hESC) carry a risk of teratoma formation by contaminating undifferentiated hESC. We generated 10 monoclonal antibodies (mAbs) against surface antigens of undifferentiated hESC, showing strong reactivity against undifferentiated, but not differentiated hESC. The mAbs did not cross react with mouse fibroblasts and showed weak to no reactivity against human embryonal carcinoma cells. Notably, one antibody (mAb 84) is cytotoxic to undifferentiated hESC and NCCIT cells in a concentration-dependent, complement-independent manner. mAb 84 induced cell death of undifferentiated, but not differentiated hESC within 30 minutes of incubation, and immunoprecipitation of the mAb-antigen complex revealed that the antigen is podocalyxin-like protein-1. Importantly, we observed absence of tumor formation when hESC and NCCIT cells were treated with mAb 84 prior to transplantation into severe combined immunodeficiency mice. Our data indicate that mAb 84 may be useful in eliminating residual hESC from differentiated cells populations for clinical applications. Disclosure of potential conflicts of interest is found at the end of this article.
Choo
Andre B
AB
Stem Cell Group, Bioprocessing Technology Institute, 20 Biopolis Way #06-01, Singapore. andre_choo@bti.a-star.edu.sg
Tan
Heng Liang
HL
Ang
Sheu Ngo
SN
Fong
Wey Jia
WJ
Chin
Angela
A
Lo
Jennifer
J
Zheng
Lu
L
Hentze
Hannes
H
Philp
Robin J
RJ
Oh
Steve K W
SK
Yap
Miranda
M
D000818:Animals; D000906:Antibodies; D000911:Antibodies, Monoclonal; D002454:Cell Differentiation; D002460:Cell Line; D002470:Cell Survival; D053595:Embryonic Stem Cells / Q000166:cytology* / Q000502:physiology; D005434:Flow Cytometry; D006367:HeLa Cells; D006801:Humans; D051379:Mice; D012795:Sialoglycoproteins / Q000032:analysis* / Q000276:immunology
D000906:Antibodies; D000911:Antibodies, Monoclonal; D012795:Sialoglycoproteins; C001643:podocalyxin