差异超速离心法对不同种属间充质干细胞外泌体分离的可靠性研究

doi:10.13418/j.issn.1001-165x.2022.3.09

Chinese Journal of Clinical Anatomy ›› 2022, Vol. 40 ›› Issue (3): 292-296.doi: 10.13418/j.issn.1001-165x.2022.3.09

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Study on the reliability of differential hypervelocity centrifugation for isolation of mesenchymal stem cells derived exosomes from different species

Zhao Renli^1,2, Lai Guohua¹, Wu Jiachang¹, Wu Mingjie¹, Zhuang Weida¹, Ouyang Jun³*, Sang Hongxun^1,2*

1. Department of Orthopedics, Shenzhen Hospital of Southern Medical University, Shenzhen 518100, China; 2. The Third Clinical Medical College of Southern Medical University, Guangzhou 510630, China; 3. Department of Human Anatomy, Guangdong Provincial Key Laboratory of Medical Biomechanics, Southern Medical University, Guangzhou 510515, China

Received:2021-05-31 Online:2022-05-25 Published:2022-05-31

Abstract

Abstract: Objective　To isolate and culture the mouse and rat bone marrow mesenchymal stem cells (BMSCs)， purify the exosome by differential hypervelocity centrifugation， identify the morphology and biological function of exosomes and analyze the reliability of this method for exosome isolation.　Methods　BMSCs of 3-5 passages were differentiated by osteogenic, adipogenic and chondrogenic induction and then stained. The surface markers of BMSCs were identified by flow cytometry. Exosomes were identified by transmission electron microscopy, nanoparticle tracer analysis and Western Blot. Results Stained after osteogenic, adipogenic and chondrogenic induction, BMSCs were proved to have the unique potential of multidirectional differentiation. Flow cytometry results showed that mouse and rat BMSCs expressed the surface antigen of stem cells. Under transmission electron microscopy, exosomes showed a typical goblet structure with an average particle size of 107 nm and 152 nm. Western Blot results showed that the exosomes expressed landmark proteins.　Conclusions　Mouse and rat BMSCs are successfully isolated and cultured. Meanwhile, differential hypervelocity centrifugation is suitable and reliable to isolate and purify exosomes from BMSCs.

Key words: Bone marrow mesenchymal stem cells, 　Exosomes;　Differential hypervelocity centrifugation

CLC Number:

Q249

Zhao Renli, Lai Guohua, Wu Jiachang, Wu Mingjie, Zhuang Weida, Ouyang Jun, Sang Hongxun. Study on the reliability of differential hypervelocity centrifugation for isolation of mesenchymal stem cells derived exosomes from different species[J]. Chinese Journal of Clinical Anatomy, 2022, 40(3): 292-296.

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Study on the reliability of differential hypervelocity centrifugation for isolation of mesenchymal stem cells derived exosomes from different species

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