AI Article Synopsis
- This study aimed to explore how the EP4A receptor influences the growth of human CD34 cells, which are important stem cells, in a lab setting.
- Researchers collected blood samples and used specific techniques to isolate these cells and determine the best conditions for their growth when treated with EP4A.
- The findings indicated that EP4A significantly increased the proliferation of CD34 cells and activated the Wnt/β-catenin signaling pathway, suggesting its potential role in stem cell therapy.
Article Abstract
Objective: To investigate the potential signaling pathway that regulates the proliferation of human CD34 cells stimulated by prostaglandin E2 receptor 4 agonist (EP4A) in vitro.
Methods: Twenty samples of peripheral blood containing stem cells were collected from the G-CSF mobilized healthy donors in our department of hematology. Human CD34 cells were isolated by magnetic activated cell sorting (MACS) microbeads kit. The Cell Counting Kit-8 (CCK8) assay was used to determine the optimal concentration and time of EP4A to promote human CD34 cell proliferation in vitro. Under the optimal condition, quantitative real-time polymerase chain reaction (qRT-PCR) was used to detect mRNA level of β-catenin, and Western blot was used to assay protein expression of β-catenin and P-GSK-3β in human CD34 cells treated with EP4A.
Results: Culturing with 10 µmol/L EP4A for 72 h, it was found that EP4A promoted human CD34 cell proliferation significantly, and the proliferation rate of human CD34 cells was 1.36 times higher than that of the control(P=0.002). Under the optimal condition, it was also found that EP4A enhanced the β-catenin expression at both mRNA and protein levels, and up-regulated phosphorylation of GSK-3β in human CD34 cells, but these effects could be inhibited by the EP4A antagonist EP4AA.
Conclusion: EP4A can enhance human CD34 cell proliferation in vitro by activating Wnt/β-catenin signaling pathway.
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| http://dx.doi.org/10.7534/j.issn.1009-2137.2017.03.004 | DOI Listing |
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