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Title 

Heat shock 70-kDa protein 8 isoform 1 is expressed on the surface of human embryonic stem cells and downregulated upon differentiation

Authors 

Yeon Sung SonJae Hyun ParkYoung Kook KangJin Sung ParkHong Seo ChoiJi Young LimJ E LeJ B LeeM S KoYong Sam KimJeong Heon KoH S YoonK W LeeR H SeongS Y MoonChun Jeih RyuHyo Jeong Hong

Publisher 

Wiley-Blackwell

Issue Date 

2005

Citation 

Stem Cells, vol. 23, no. 10, pp. 1502-1513

Keywords 

cell-surface markerheat shock protein 70human embryonic stem cellscell surface markerheat shock proteinheat shock protein 8 isoform 1immunoglobulin G1protein tra[1-60]protein tra[1-81]animal cell

Abstract 

The cell-surface markers used routinely to define the undifferentiated state and pluripotency of human embryonic stem cells (hESCs) are those used in mouse embryonic stem cells (mESCs) because of a lack of markers directly originated from hESC itself. To identify more hESC-specific cell-surface markers, we generated a panel of monoclonal antibodies (MAbs) by immunizing the irradiated cell clumps of hESC line Miz-hES1, and selected 26 MAbs that were able to bind to Miz-hES1 cells but not to mESCs, mouse embryonic fibroblast cells, and STO cells. Most antibodies did not bind to human neural progenitor cells derived from the Miz-hES1 cells, either. of these, MAb 20-202S (IgG1, κ) immunoprecipitated a cell-surface protein of 72-kDa from the lysate of biotin-labeled Miz-hES1 cells, which was identified to be heat shock 70-kDa protein S isoform 1 (HSPA8) by quadrupole time-of-flight tandem mass spectrometry. Immunocytochemical analyses proved that the HSPA8 protein was also present on the surface of hESC lines Miz-hES4, Miz-hES6, and HSF6. Two-color flow cytometric analysis of Miz-hES1 and HSF6 showed the coexpression of the HSPA8 protein with other hESC markers such as stage-specific embryonic antigen 3 (SSEA3), SSEA4, TRA-1-60, and TRA-1-81. Flow cytometric and Western blot analyses using various cells showed that MAb 20-202S specifically bound to the HSPA8 protein on the surface of Miz-hES1, contrary to other anti-HSP70 antibodies examined. Furthermore, the surface expression of the HSPA8 protein on Miz-hES1 was markedly downregulated upon differentiation. These data indicate that a novel MAb 20-202S recognizes the HSPA8 protein on the surface of hESCs and suggest that the HSPA8 protein is a putative cell-surface marker for undifferentiated hESCs.

ISSN 

1066-5099

Link 

http://dx.doi.org/10.1634/stemcells.2004-0307

Appears in Collections

1. Journal Articles > Journal Articles

Registered Date

2019-05-02


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