PMID- 33148384
OWN - NLM
STAT- MEDLINE
DCOM- 20201106
LR  - 20201106
IS  - 1007-8738 (Print)
IS  - 1007-8738 (Linking)
VI  - 36
IP  - 10
DP  - 2020 Oct
TI  - [Comparison of four methods for extracting extracellular vesicles from human 
      umbilical cord mesenchymal stem cells].
PG  - 897-902
AB  - Objective To compare the efficiency of four methods for extracting extracellular 
      vesicles (EVs) from human umbilical cord mesenchymal stem cells(hUCMSCs). Methods 
      EVs were isolated from the conditioned medium of hUCMSCs by ultracentrifugation 
      (group A), or ultrafiltration combined with ultracentrifugation (group B), or 
      ultrafiltration combined with polyethylene glycol precipitation (group C), or 
      ultrafiltration combined with aqueous two phase system (group D). The total 
      protein concentration of EVs in each group was determined by BCA method. The 
      expression of Alix, CD9, and calnexin were detected by Western blotting. The 
      morphology of EVs was analyzed by transmission electron microscopy. The particle 
      size distribution and particle concentration of EVs were measured by nanoparticle 
      tracking analysis. Results The total protein concentrations of EVs extracted by 
      the above four methods were (1.92+/-1.77) mug/muL, (18.1+/-1.07) mug/muL, (6.33+/-1.02) 
      mug/muL, (36.48+/-23.13) mug/muL from group A to D respectively. We observed the 
      expression of CD9 and Alix, but not calnexin, in EVs from group A, B and C. 
      However, the expression levels of CD9 and Alix were lowest in group C. In 
      addition, the expression of CD9, Alix and calnexin were undetectable in EVs from 
      group D. The particle concentrations of EVs in group A, B and C were 0.85x10(11) 
      particles/mL, 0.63x10(11) particles/mL, 1.83x10(11) particles/mL, respectively. 
      Meanwhile, the particle distributions were all within the size range of EVs. We 
      also observed the typical saucer-like membrane structure in EVs from group A, B 
      and C. Conclusion The method of ultrafiltration combined with ultracentrifugation 
      could be applied to the experiments demanding large amounts of EVs. The method of 
      ultracentrifugation is recommended for the extraction of little amounts of EVs 
      due to the lower risk of EV fragmentation.
FAU - He, Jing
AU  - He J
AD  - Department of Biochemistry and Molecular Biology, School of Basic Medical 
      Sciences, Wuhan University, Wuhan 430072, China.
FAU - Deng, Taoran
AU  - Deng T
AD  - Reproductive Medicine Center, Tongji Hospital, Tongji Medical College, Huazhong 
      University of Science and Technology, Wuhan 430075, China.
FAU - Li, Changyong
AU  - Li C
AD  - Department of Biochemistry and Molecular Biology, School of Basic Medical 
      Sciences, Wuhan University, Wuhan 430072, China.
FAU - Li, Yufeng
AU  - Li Y
AD  - Reproductive Medicine Center, Tongji Hospital, Tongji Medical College, Huazhong 
      University of Science and Technology, Wuhan 430075, China.
FAU - Wu, Dongcheng
AU  - Wu D
AD  - Department of Biochemistry and Molecular Biology, School of Basic Medical 
      Sciences, Wuhan University, Wuhan 430072; Hamilton Biotechnology-Co., LTD, Wuhan 
      430075, China. *Corresponding authors, E-mail: bcdcwu@hotmail.com.
FAU - Wu, Mingfu
AU  - Wu M
AD  - Department of Obsterics and Gynecology, Tongji Hospital, Tongji Medical College, 
      Huazhong University of Science and Technology, Wuhan 430030, China. 
      *Corresponding authors, E-mail: mingfuwutj@163.com.
LA  - chi
PT  - Comparative Study
PT  - Journal Article
PL  - China
TA  - Xi Bao Yu Fen Zi Mian Yi Xue Za Zhi
JT  - Xi bao yu fen zi mian yi xue za zhi = Chinese journal of cellular and molecular 
      immunology
JID - 101139110
RN  - 0 (Culture Media, Conditioned)
SB  - IM
MH  - *Culture Media, Conditioned
MH  - *Extracellular Vesicles
MH  - Humans
MH  - *Mesenchymal Stem Cells
MH  - Microscopy, Electron, Transmission
MH  - Ultracentrifugation
MH  - Ultrafiltration
MH  - Umbilical Cord/cytology
EDAT- 2020/11/06 06:00
MHDA- 2020/11/11 06:00
CRDT- 2020/11/05 05:37
PHST- 2020/11/05 05:37 [entrez]
PHST- 2020/11/06 06:00 [pubmed]
PHST- 2020/11/11 06:00 [medline]
PST - ppublish
SO  - Xi Bao Yu Fen Zi Mian Yi Xue Za Zhi. 2020 Oct;36(10):897-902.