Construction of Cell Model to CD47 Target Research
MEI Endian1, SHEN Junjie2, CHEN Xuejiao2, QIAN Cheng1*
CD47 is a new immune checkpoint. However, CD47 targeted studies lack reliable research models because CD47 widely expressed in human-derived cells and CD47-negative human-derived cells haven’t been found. In this study, taking CHO (Chinese hamster ovary) cell to be a basis, human-derived CD47 lentiviral vector was prepared and CHO was infected to construct CHO-CD47 cell line. The CHO-CD47 was evaluated through flow cytometry, RT-PCR (reverse transcription-polymerase chain reaction) and biological binding detections. The result of flow cytometry detection demonstrated that CHO-CD47 cell line expressed human-derived CD47 and the positive rate was 94.7%, which kept stable (96.4%) after freeze-thawing and passaging 10 times. The result of RNA level detection demonstrated that CHO-CD47 expressed human-derived CD47 mRNA. In addition, the result of protein level detection showed that CHO-CD47 could bind to human-derived CD47 ligands SIRPα (signal regulatory protein α) and SIRPγ (signal regulatory protein γ) recombinant protein successfully. In summary, the construction of the CHO-CD47 cell model was successful. It could be used to verify the binding ability of CD47 target studies, providing a reliable research model for CD47 targeted studies and a certain research basis for cancer therapy studies targeting CD47.
CN
EN