Human ocular epithelial cells endogenously expressing SOX2 and OCT4 yield high efficiency of pluripotency reprogramming

Ming Wai Poon, Jia He, Xiaowei Fang, Zhao Zhang, Weixin Wang, Junwen Wang, Fangfang Qiu, Hung Fat Tse, Wei Li, Zuguo Liu, Qizhou Lian

Research output: Contribution to journalArticle

7 Citations (Scopus)

Abstract

A variety of pluripotency reprogramming frequencies from different somatic cells has been observed, indicating cell origin is a critical contributor for efficiency of pluripotency reprogramming. Identifying the cell sources for efficient induced pluripotent stem cells (iPSCs) generation, and defining its advantages or disadvantages on reprogramming, is therefore important. Human ocular tissue-derived conjunctival epithelial cells (OECs) exhibited endogenous expression of reprogramming factors OCT4A (the specific OCT 4 isoform on pluripotency reprogramming) and SOX2. We therefore determined whether OECs could be used for high efficiency of iPSCs generation. We compared the endogenous expression levels of four pluripotency factors and the pluripotency reprograming efficiency of human OECs with that of ocular stromal cells (OSCs). Real-time PCR, microarray analysis, Western blotting and immunostaining assays were employed to compare OECiPSCs with OSCiPSCs on molecular bases of reprogramming efficiency and preferred lineage-differentiation potential. Using the traditional KMOS (KLF4, C-MYC, OCT4 and SOX2) reprogramming protocol, we confirmed that OECs, endogenously expressing reprogramming factors OCT4A and SOX2, yield very high efficiency of iPSCs generation (∼1.5%). Furthermore, higher efficiency of retinal pigmented epithelial differentiation (RPE cells) was observed in OECiPSCs compared to OSCiPSCs or skin fibroblast iMR90iPSCs. The findings in this study suggest that conjunctival-derived epithelial (OECs) cells can be easier converted to iPSCs than conjunctival-derived stromal cells (OSCs). This cell type may also have advantages in retinal pigmented epithelial differentiation.

Original languageEnglish (US)
Article numbere0131288
JournalPLoS One
Volume10
Issue number7
DOIs
StatePublished - Jul 1 2015
Externally publishedYes

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Induced Pluripotent Stem Cells
stromal cells
epithelial cells
Stromal Cells
eyes
Epithelial Cells
Stem cells
cells
Microarray Analysis
somatic cells
skin (animal)
cell differentiation
fibroblasts
Real-Time Polymerase Chain Reaction
Cell Differentiation
Protein Isoforms
quantitative polymerase chain reaction
Western blotting
Fibroblasts
Western Blotting

ASJC Scopus subject areas

  • Agricultural and Biological Sciences(all)
  • Biochemistry, Genetics and Molecular Biology(all)
  • Medicine(all)

Cite this

Human ocular epithelial cells endogenously expressing SOX2 and OCT4 yield high efficiency of pluripotency reprogramming. / Poon, Ming Wai; He, Jia; Fang, Xiaowei; Zhang, Zhao; Wang, Weixin; Wang, Junwen; Qiu, Fangfang; Tse, Hung Fat; Li, Wei; Liu, Zuguo; Lian, Qizhou.

In: PLoS One, Vol. 10, No. 7, e0131288, 01.07.2015.

Research output: Contribution to journalArticle

Poon, MW, He, J, Fang, X, Zhang, Z, Wang, W, Wang, J, Qiu, F, Tse, HF, Li, W, Liu, Z & Lian, Q 2015, 'Human ocular epithelial cells endogenously expressing SOX2 and OCT4 yield high efficiency of pluripotency reprogramming', PLoS One, vol. 10, no. 7, e0131288. https://doi.org/10.1371/journal.pone.0131288
Poon, Ming Wai ; He, Jia ; Fang, Xiaowei ; Zhang, Zhao ; Wang, Weixin ; Wang, Junwen ; Qiu, Fangfang ; Tse, Hung Fat ; Li, Wei ; Liu, Zuguo ; Lian, Qizhou. / Human ocular epithelial cells endogenously expressing SOX2 and OCT4 yield high efficiency of pluripotency reprogramming. In: PLoS One. 2015 ; Vol. 10, No. 7.
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