Preparation of monoclonal antibodies against human mesenchymal stem cells.
- Author:
Zhi-xiong LIAO
1
;
Jian-gen SHEN
;
Xiao-yu LAI
Author Information
- Publication Type:Journal Article
- MeSH: Animals; Antibodies, Monoclonal; biosynthesis; Antibody Specificity; Bone Marrow Cells; cytology; immunology; Fluorescent Antibody Technique; methods; HL-60 Cells; Humans; Hybridomas; secretion; Immunoglobulin G; immunology; Immunoglobulin M; immunology; Mesenchymal Stromal Cells; cytology; immunology; Mice; Mice, Inbred BALB C; Rats
- From: Journal of Zhejiang University. Medical sciences 2006;35(5):517-522
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVETo prepare monoclonal antibodies (McAbs) against human mesenchymal stem cells (hMSCs) and to study their biological characteristics.
METHODSBALB/C mice were immunized with pooled hMSCs. McAbs were prepared by hybridoma technique and their biological characteristics were analyzed by indirect immunofluorescence, immunohistochemistry and flow cytometry.
RESULTFive hybridoma cell lines were successfully established, which secret McAbs specifically against hMSCs. Investigations showed that all these McAb reacted only to hMSCs and had no cross-reaction to other human cells, the relative affinities of 5 McAbs were 1x10(6) (ZUB1), 1x10(5) (ZUB4), 1x10(6) (ZUC3), ZUE12 (1x10(5)) and 1x10(5) (ZUF10), respectively. Isotype analysis showed that ZUB1, ZUE12, ZUF10 against the same isotype, while ZUC3, ZUB4 against other two different isotypes alone. Flow cytometric analysis showed that the positive expression rate of cultured hMSCs was 87.39% (ZUB1), 88.07% (ZUB4), 88.12% (ZUC3), 69.89% (ZUE12) and 83.67% (ZUF10).
CONCLUSIONThe prepared five McAbs can specifically react against hMSCs, which can be used for selection and study of hMCSs.
