Tumstatin Transgenic Megakaryocyte Produce Anti-angiogenesis Platelet In NOD/SCID Mice

Objective Clinical chemotherapy induced thrombocytopenia, but platelet transfusion directly would increase the risk of tumor cell growth and metastasis. This study is about the tumstatin gene combined with hematopoietic stem cell (HSC) CD34+ cells in vitro generating transgenic megakaryocytes (MKS), then infused to NOD/SCID mice, try to understand the antitumor effect of tumstatin transgenic MKs in NOD/SCID mice in vivo. In previous study, we have successfully prepared transgenic MKs, this experiment is mainly about infused transgenic MKs to NOD/SCID mice to investigate tumstatin transgenic megakaryocyte producing platelets in NOD/SCID mice and its anti-angiogenesis fuction.. This study provide basis for the treatment of tumors with tumstatin transgenic platelet.Methods (1)After identification in vitro, tumstatin transgenic megakaryocyte were injected into NOD/SCID mice. Analysed tumstatin transgenic megakaryocyte with Wright’s stain, and them, which were randomly divided into transgenic group, the non-transgenic group and the control group. (2)Mice peripheral blood were regularly taken, flow cytometry was used to test transgenic platelets production in NOD/SCID mice; (3)Immunofluorescence was used to test the expression of tumstatin; (4)Endothelial cells tubular structure formation test was used to test anti-angiogenesis fuction; (5)Platelet aggregation test was used to test aggregation function.Results(1)Morphological analysis showed that tumstatin transgenic cells showed typical morphology of megakaryocyte; (2)After 3 d of transgenic megakaryocyte infusion, human platelets can be detected in peripheral blood of NOD/SCID mice by flow cytometry; (3) vWF protein and tumstatin protein expression were observed simultaneously in part of peripheral blood platelets by immunofluorescence; (4)Transgenic platelet can obviously inhibit the endothelial cells tubular structure formation, proved by endothelial cells tubular structure formation test; (5)Transgenic platelet can keep nomal aggregation function, proved by platelet aggregation test.Conclusion Experiment results show that tumstatin transgenic megakaryocyte were succesfully infused into NOD/SCID mice, and can produce anti-angiogenesis platelets having normal aggregation function in NOD/SCID mice, which lay the foundation for further anti-tumor research.