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  • Role of CD133 in human embryonic stem cell proliferation and teratoma formation.

Role of CD133 in human embryonic stem cell proliferation and teratoma formation.

Stem cell research & therapy (2020-05-29)
Hua Wang, Peng Gong, Jie Li, Yudong Fu, Zhongcheng Zhou, Lin Liu
ABSTRACT

Pluripotent stem cells (PSCs), including human embryonic stem cells (hESCs), hold great potential for regenerative medicine and cell therapy. One of the major hurdles hindering the clinical development of PSC-based therapy is the potential risk of tumorigenesis. CD133 (Prominin 1, PROM1) is a transmembrane protein whose mRNA and glycosylated forms are highly expressed in many human cancer cell types. CD133 also serves as a cancer stem cell (CSC) marker associated with cancer progression and patient outcome. Interestingly, CD133 is highly expressed in hESCs as well as in human preimplantation embryos, but its function in hESCs has remained largely unknown. CD133 knockout hESC WA26 cell line was generated with CRISPR/Cas9. CD133 knockout and wide type hESC lines were subjected to pluripotency, proliferation, telomere biology, and teratoma tests; the related global changes and underlying mechanisms were further systemically analyzed by RNA-seq. CD133 deficiency did not affect hESC pluripotency or in vivo differentiation into three germ layers but significantly decreased cell proliferation. RNA-seq revealed that CD133 deficiency dysregulated the p53, PI3K-Akt, AMPK, and Wnt signaling pathways. Alterations in these pathways have been implicated in tumor proliferation and apoptotic escape. Our data imply that CD133 could be an additional target and used as a selective marker to sort and eliminate undifferentiated cells in reducing potential teratoma formation risk of hESCs in regenerative medicine.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
Anti-Stage-Specific Embryonic Antigen-4 Antibody, clone MC-813-70, clone MC-813-70, Chemicon®, from mouse
Sigma-Aldrich
Anti-TRA-1-81 Antibody, clone TRA-1-81, clone TRA-1-81, Chemicon®, from mouse
Sigma-Aldrich
Anti-Sox2 Antibody, Chemicon®, from rabbit
Roche
TeloTAGGG Telomere Length Assay, sufficient for ≤50 reactions, kit of 1 (15 components), suitable for cell culture