Establishment and Identification of a CiPSC Lineage Reprogrammed from FSP-tdTomato Mouse Embryonic Fibroblasts (MEFs)

Safety issues associated with transcription factors or viruses may be avoided with the use of chemically induced pluripotent stem cells (CiPSCs), thus promoting their clinical application. Previously, we had successfully developed and standardized an induction method using small-molecule compound, w...

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Bibliographic Details
Published in:Stem cells international 2018-01, Vol.2018 (2018), p.1-8
Main Authors: Yang, Bin, Zhao, Mingyi, Zhu, Ping, Yu, Shengyong, Zhou, Chunhua, Zhou, Wenyi, Wu, Chuman, Qin, Yue, Cai, Baomei, Xie, Wenxiu, Chen, Ruiping, Kuang, Junqi
Format: Article
Language:English
Subjects:
Biology
Cardiomyocytes
Cell lineage
Differentiation (biology)
Efficiency
Embryo cells
Embryo fibroblasts
Epigenetics
Fibroblasts
Fluorescence
Gene expression
Nucleoli
Organic chemistry
Pluripotency
Proteins
Skin
Stem cell transplantation
Stem cells
Teratoma
Tracking systems
Transcription factors
Transplantation
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Summary:Safety issues associated with transcription factors or viruses may be avoided with the use of chemically induced pluripotent stem cells (CiPSCs), thus promoting their clinical application. Previously, we had successfully developed and standardized an induction method using small-molecule compound, with simple operation, uniform induction conditions, and clear constituents. In order to verify that the CiPSCs were indeed reprogrammed from mouse embryonic fibroblasts (MEFs), and further explore the underlying mechanisms, FSP-tdTomato mice were used to construct a fluorescent protein-tracking system of MEFs, for revealing the process of CiPSC reprogramming. CiPSCs were identified by morphological analysis, mRNA, and protein expression of pluripotency genes, as well as teratoma formation experiments. Results showed that after 40-day treatment of tdTomato-MEFs with small-molecule compounds, the cells were presented with prominent nucleoli, high core-to-cytoplasmic ratio, round shape, group and mass arrangement, and high expression of pluripotency gene. These cells could differentiate into three germ layer tissues in vivo. As indicated by the above results, tdTomato-MEFs could be reprogrammed into CiPSCs, a lineage that possesses pluripotency similar to mouse embryonic stem cells (mESCs), with the use of small-molecule compounds. The establishment of CiPSC lineage, tracked by fluorescent protein, would benefit further studies exploring its underlying mechanisms. With continuous expression of fluorescent proteins during cellular differentiation, this cell lineage could be used for tracking CiPSC transplantation and differentiation into functional cells.
ISSN:1687-966X
1687-9678
DOI:10.1155/2018/5965727