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Chang, Young Tae
복잡계 자기조립 연구단
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Novel live cell fluorescent probe for human-induced pluripotent stem cells highlights early reprogramming population

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Title
Novel live cell fluorescent probe for human-induced pluripotent stem cells highlights early reprogramming population
Author(s)
Sriram, Sandhya; Kang, Nam-Young; Subramanian, Subha; Nandi, Tannistha; Sudhagar, Samydurai; Xing, Qiaorui; Tong, Gerine Jin-Ling; Chen, Allen Kuan-Liang; Srijaya, Thekkeparambil Chandrabose; Tan, Patrick; Loh, Yuin-Han; Young-Tae Chang; Sugii, Shigeki
Publication Date
2021-02
Journal
STEM CELL RESEARCH & THERAPY, v.12, no.1
Publisher
BMC
Abstract
Background Despite recent rapid progress in method development and biological understanding of induced pluripotent stem (iPS) cells, there has been a relative shortage of tools that monitor the early reprogramming process into human iPS cells. Methods We screened the in-house built fluorescent library compounds that specifically bind human iPS cells. After tertiary screening, the selected probe was analyzed for its ability to detect reprogramming cells in the time-dependent manner using high-content imaging analysis. The probe was compared with conventional dyes in different reprogramming methods, cell types, and cell culture conditions. Cell sorting was performed with the fluorescent probe to analyze the early reprogramming cells for their pluripotent characteristics and genome-wide gene expression signatures by RNA-seq. Finally, the candidate reprogramming factor identified was investigated for its ability to modulate reprogramming efficiency. Results We identified a novel BODIPY-derived fluorescent probe, BDL-E5, which detects live human iPS cells at the early reprogramming stage. BDL-E5 can recognize authentic reprogramming cells around 7 days before iPS colonies are formed and stained positive with conventional pluripotent markers. Cell sorting of reprogrammed cells with BDL-E5 allowed generation of an increased number and higher quality of iPS cells. RNA sequencing analysis of BDL-E5-positive versus negative cells revealed early reprogramming patterns of gene expression, which notably included CREB1. Reprogramming efficiency was significantly increased by overexpression of CREB1 and decreased by knockdown of CREB1. Conclusion Collectively, BDL-E5 offers a valuable tool for delineating the early reprogramming pathway and clinically applicable commercial production of human iPS cells.
URI
https://pr.ibs.re.kr/handle/8788114/9597
DOI
10.1186/s13287-021-02171-6
ISSN
1757-6512
Appears in Collections:
Center for Self-assembly and Complexity(복잡계 자기조립 연구단) > 1. Journal Papers (저널논문)
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