Comparison of Ex Vivo Expanded and Highly Purified NK Cell-Mediated Cytotoxicity Detected by 3 Different Staining Methods of Flow Cytometry
OBJECTIVE: To compare the cytotoxicity of ex vivo expanded NK cells detected by flow cytometry with 3 different staining methods.
METHODS: NK cells were collected from peripheral blood on the 17th day after culture. The cultured cells were divided into 3 groups: group A , B, and C. The cells in group A were stained with CFSE/Annectin-V/7-AAD; the cells in group B were stained with Annectin-V/PI, and the cells in group C cells were stained with CFSE/PI. The E:T ratios in 3 groups were 10:1, 20:1 and 40:1, respectively, the K562 cells were incubated with NK cells for 4 hrs.
RESULTS: The purity of NK cells(CD3-CD56+) reached to (16.34±10.51)% on day 0 and to (83.63±10.63)% on the day 17 after incubation(P<0.05); the cytotoxicity of group A was significantly higher than thay of group B at different E:T ratio (P<0.05). The cytotoxicities in A, B, C groups at E:T ratio=10:1 were (36.56±3.69)%, (10.85±2.09)% and (22.35±2.71)% respectively; the cytotoxicities in A, B, C groups at E:T ratio=9:1 were (47.83±5.52)%, (39.07±5.55)% and (29.61±4.81)%; the cytotoxicities in A, B, C groups at E:T ratio=40:1 were (67.7±4.77)%, (51.51±4.43)% and (44.12±5.62)% respectively. Meanwhile, the cytotoxicity in group A was significantly higher than that in group C at different E:T ratio (P<0.05), the percentage of cytotoxicity was (36.56±3.69)% vs (10.85±2.09)%, (47.83±5.52)% vs (29.61±4.81)%, (67.7±4.77)% vs (44.12±5.62)%, respectively.
CONCLUSION: CFSE/Annectin-V/7-AAD is able to clearly show human NK cell cytotoxicity against human tumors. Moreover, this staining technique also allows to distinguish different stages of cytotoxic killing as early and late apoptotic phase.
| Medienart: |
E-Artikel |
|---|
| Erscheinungsjahr: |
2016 |
|---|---|
| Erschienen: |
2016 |
| Enthalten in: |
Zur Gesamtaufnahme - volume:24 |
|---|---|
| Enthalten in: |
Zhongguo shi yan xue ye xue za zhi - 24(2016), 6 vom: 27. Dez., Seite 1691-1697 |
| Sprache: |
Chinesisch |
|---|
| Beteiligte Personen: |
Guo, Ji-Qiang [VerfasserIn] |
|---|
| Links: |
|---|
| Themen: |
|---|
| Anmerkungen: |
Date Completed 28.06.2018 Date Revised 23.10.2018 published: Print Citation Status MEDLINE |
|---|
| doi: |
10.7534/j.issn.1009-2137.2016.06.014 |
|---|
| funding: |
|
|---|---|
| Förderinstitution / Projekttitel: |
|
| PPN (Katalog-ID): |
NLM267479972 |
|---|
| LEADER | 01000naa a22002652 4500 | ||
|---|---|---|---|
| 001 | NLM267479972 | ||
| 003 | DE-627 | ||
| 005 | 20231224220755.0 | ||
| 007 | cr uuu---uuuuu | ||
| 008 | 231224s2016 xx |||||o 00| ||chi c | ||
| 024 | 7 | |a 10.7534/j.issn.1009-2137.2016.06.014 |2 doi | |
| 028 | 5 | 2 | |a pubmed24n0891.xml |
| 035 | |a (DE-627)NLM267479972 | ||
| 035 | |a (NLM)28024478 | ||
| 040 | |a DE-627 |b ger |c DE-627 |e rakwb | ||
| 041 | |a chi | ||
| 100 | 1 | |a Guo, Ji-Qiang |e verfasserin |4 aut | |
| 245 | 1 | 0 | |a Comparison of Ex Vivo Expanded and Highly Purified NK Cell-Mediated Cytotoxicity Detected by 3 Different Staining Methods of Flow Cytometry |
| 264 | 1 | |c 2016 | |
| 336 | |a Text |b txt |2 rdacontent | ||
| 337 | |a ƒaComputermedien |b c |2 rdamedia | ||
| 338 | |a ƒa Online-Ressource |b cr |2 rdacarrier | ||
| 500 | |a Date Completed 28.06.2018 | ||
| 500 | |a Date Revised 23.10.2018 | ||
| 500 | |a published: Print | ||
| 500 | |a Citation Status MEDLINE | ||
| 520 | |a OBJECTIVE: To compare the cytotoxicity of ex vivo expanded NK cells detected by flow cytometry with 3 different staining methods | ||
| 520 | |a METHODS: NK cells were collected from peripheral blood on the 17th day after culture. The cultured cells were divided into 3 groups: group A , B, and C. The cells in group A were stained with CFSE/Annectin-V/7-AAD; the cells in group B were stained with Annectin-V/PI, and the cells in group C cells were stained with CFSE/PI. The E:T ratios in 3 groups were 10:1, 20:1 and 40:1, respectively, the K562 cells were incubated with NK cells for 4 hrs | ||
| 520 | |a RESULTS: The purity of NK cells(CD3-CD56+) reached to (16.34±10.51)% on day 0 and to (83.63±10.63)% on the day 17 after incubation(P<0.05); the cytotoxicity of group A was significantly higher than thay of group B at different E:T ratio (P<0.05). The cytotoxicities in A, B, C groups at E:T ratio=10:1 were (36.56±3.69)%, (10.85±2.09)% and (22.35±2.71)% respectively; the cytotoxicities in A, B, C groups at E:T ratio=9:1 were (47.83±5.52)%, (39.07±5.55)% and (29.61±4.81)%; the cytotoxicities in A, B, C groups at E:T ratio=40:1 were (67.7±4.77)%, (51.51±4.43)% and (44.12±5.62)% respectively. Meanwhile, the cytotoxicity in group A was significantly higher than that in group C at different E:T ratio (P<0.05), the percentage of cytotoxicity was (36.56±3.69)% vs (10.85±2.09)%, (47.83±5.52)% vs (29.61±4.81)%, (67.7±4.77)% vs (44.12±5.62)%, respectively | ||
| 520 | |a CONCLUSION: CFSE/Annectin-V/7-AAD is able to clearly show human NK cell cytotoxicity against human tumors. Moreover, this staining technique also allows to distinguish different stages of cytotoxic killing as early and late apoptotic phase | ||
| 650 | 4 | |a Journal Article | |
| 700 | 1 | |a Han, Ya-Ping |e verfasserin |4 aut | |
| 700 | 1 | |a Li, Fang |e verfasserin |4 aut | |
| 700 | 1 | |a Zhao, Qing |e verfasserin |4 aut | |
| 700 | 1 | |a Jia, Yuan |e verfasserin |4 aut | |
| 700 | 1 | |a Jin, Bao-Li |e verfasserin |4 aut | |
| 700 | 1 | |a Zhang, Jun-Ping |e verfasserin |4 aut | |
| 773 | 0 | 8 | |i Enthalten in |t Zhongguo shi yan xue ye xue za zhi |d 2000 |g 24(2016), 6 vom: 27. Dez., Seite 1691-1697 |w (DE-627)NLM122963113 |x 1009-2137 |7 nnns |
| 773 | 1 | 8 | |g volume:24 |g year:2016 |g number:6 |g day:27 |g month:12 |g pages:1691-1697 |
| 856 | 4 | 0 | |u http://dx.doi.org/10.7534/j.issn.1009-2137.2016.06.014 |3 Volltext |
| 912 | |a GBV_USEFLAG_A | ||
| 912 | |a GBV_NLM | ||
| 951 | |a AR | ||
| 952 | |d 24 |j 2016 |e 6 |b 27 |c 12 |h 1691-1697 | ||