Female germline stem cells (FGSCs) have been successfully isolated and characterized from postnatal mammalian and human ovarian tissues. However, the effects and mechanisms of action of natural small-molecule compounds on FGSCs are largely unknown. Here, we found that daidzein promoted the viability and proliferation of FGSCs. To elucidate the mechanism underlying this, we performed RNA-Sequence in daidzein-treated FGSCs and controls. The results showed that there were 153 upregulated and 156 downregulated genes in daidzein treatment. We confirmed the expression of some genes related to cell proliferation in the sequencing results by RT-PCR, such as Type C lectin domain family 11 member a (Clec11a), Mucin1 (Muc1), Glutathione peroxidase 3 (Gpx3), and Tet methylcytosine dioxygenase 1 (Tet1). The high expression of Clec11a at the protein level after daidzein treatment was also confirmed by western blotting. Furthermore, recombinant mouse Clec11a (rmClec11a) protein was shown to promote the viability and proliferation of FGSCs. However, knockdown of Clec11a inhibited the viability and proliferation of FGSCs, which could not be rescued by the administration of daidzein. These results indicate that daidzein promoted the viability and proliferation of FGSCs through Clec11a. In addition, both daidzein and rmClec11a activated the Akt signaling pathway in FGSCs. However, Clec11a knockdown inhibited this pathway, which could not be rescued by daidzein administration. Taken together, our findings revealed that daidzein activates the Akt signaling pathway to promote cell viability and proliferation through upregulating Clec11a. This study should deepen our understanding of the developmental mechanism of FGSCs and female infertility.
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Availability of Data and Material Raw sequencing reads were obtained from Gene Expression Omnibus and processed with the parameters detailed below: GSE199120.
Female germline stem cell
Type C lectin domain family 11 member a
Reverse-transcriptionpolymerase chain reaction
Quantitative real-time polymerase chain reaction
AKT serine/threonine kinase
Cell Counting Kit-8
Glutathione peroxidase 3
Tet methylcytosine dioxygenase 1
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This study was supported by National Nature Science Foundation of China (81720108017), the National Major Scientific Instruments and Equipment Development Project, National Natural Science Foundation of China (61827814).
All procedures involving animals were approved by the Institutional Animal Care and use committee of Shanghai and conducted in accordance with the National Research Council Guide for the Care and Use of Laboratory Animals.
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Li, F., Hu, X. & Wu, J. Daidzein Activates Akt Pathway to Promote the Proliferation of Female Germline Stem Cells through Upregulating Clec11a. Stem Cell Rev and Rep 18, 3021–3032 (2022). https://doi.org/10.1007/s12015-022-10394-0