PMID- 28851172
OWN - NLM
STAT- MEDLINE
DCOM- 20181214
LR  - 20181214
IS  - 0529-567X (Print)
IS  - 0529-567X (Linking)
VI  - 52
IP  - 8
DP  - 2017 Aug 25
TI  - [Experiment research of natural killer cells amplification in vitro and the 
      killing effect on ovarian cancer cells].
PG  - 545-550
LID - 10.3760/cma.j.issn.0529-567X.2017.08.008 [doi]
AB  - Objective: To amplify natural killer (NK) cells in vitro and explore its killing 
      effect on ovarian cancer cells. Methods: (1) The separation of NK cells and 
      identification. A total of 20 ml peripheral blood of one healthy volunteer was 
      collected in Nov. 2015, Peking University People's Hospital. The peripheral blood 
      mononuclear cells of normal volunteers were isolated, cultured in vitro and 
      amplificated cultivation for 14 days with K562 cells transfected and expressing 
      interleukin 21 (IL-21-K562) as nourish cells. The number and dynamic state of the 
      growth cells were monitored during the cultured process. Cells were harvested and 
      counted after 14 days cultured. The NK cells phenotypes were detected by flow 
      cytometry. (2) The killing effect of NK cells on ovarian cancer cells: the ratio 
      of effector cells (NK cells) and target cells (ovarian cancer cells and its 
      control) was 50ratio1, 20ratio1, 10ratio1, 5ratio1 or 1ratio1, NK cells killing effect on ovarian 
      cancer cells was detected by the lactate dehydrogenase (LDH) release experiments. 
      Results: (1) The results of NK cells establishment and phenotypic 
      characterization: the cells were induced in vitro for 14 days by amplification 
      culture. With the extension of incubation time, the number of NK cells increased 
      constantly, from 2.0x10(7) on day 0 to 5.1x10(9) on day 14. Obvious amplification 
      of the total number of cells were detected for 255 times. Living cells unstained 
      by trypan blue eventually reached 95% above. Before and after the induction and 
      amplification in vitro, the percentage of NK cells(CD(3)(-)CD(5)(6+)cells) in 
      CD(3)- cells were 2.33% and 85.32%, respectively (P<0.01), which covered the 
      whole lymphocytes 1.06% and 69.42%, respectively (P<0.01), which showed that NK 
      was the main cell type in the amplificated lymphocytes. (2) The killing rate of 
      NK cells on ovarian cancer cells in vitro: the results detected by LDH release 
      experiments showed that NK cells could performed strong nonspecific killing 
      effect on ovarian cancer cell lines SKOV3, HOC1A, 3AO and CAOV3, as well the 
      normal ovarian cell line T29 and NK sensitive cell line K562, and the killing 
      effect increased significantly along with the increase of effector cells and 
      target cells ratio (P<0.01). When the ratio was 1ratio1, the killing rate was 37% for 
      K562, while the rate of killing of other cells was around 10% (P<0.05). When the 
      effect-target ratio was 20ratio1 and 50ratio1, in addition to CAOV3 cells (more than 
      70%), NK cells had a kill rate of more than 80% for other ovarian cancer cells 
      lines and their control cell K562 and T29 cells (P>0.05). Conclusion: NK cells 
      could be established in vitro and have a good non-specific killing effect on 
      ovarian cancer cells.
FAU - Cheng, H Y
AU  - Cheng HY
AD  - Gynecology Oncology Center, Peking University People's Hospital, Beijing 100044, 
      China.
FAU - Ye, X
AU  - Ye X
FAU - Ma, R Q
AU  - Ma RQ
FAU - Chang, X H
AU  - Chang XH
FAU - Cui, H
AU  - Cui H
LA  - chi
PT  - Journal Article
PL  - China
TA  - Zhonghua Fu Chan Ke Za Zhi
JT  - Zhonghua fu chan ke za zhi
JID - 16210370R
RN  - 0 (Interleukins)
RN  - MKM3CA6LT1 (interleukin-21)
SB  - IM
MH  - Carcinoma, Ovarian Epithelial/drug therapy/metabolism
MH  - Cell Proliferation
MH  - Female
MH  - Flow Cytometry
MH  - Humans
MH  - Interleukins/metabolism
MH  - K562 Cells
MH  - Killer Cells, Natural/*drug effects/*metabolism
MH  - Leukocytes, Mononuclear
MH  - *Ovarian Neoplasms/drug therapy/metabolism
OTO - NOTNLM
OT  - Immunotherapy, adoptive
OT  - Killer cells, natural
OT  - Ovarian neoplasms
EDAT- 2017/08/31 06:00
MHDA- 2018/12/15 06:00
CRDT- 2017/08/31 06:00
PHST- 2017/08/31 06:00 [entrez]
PHST- 2017/08/31 06:00 [pubmed]
PHST- 2018/12/15 06:00 [medline]
AID - 10.3760/cma.j.issn.0529-567X.2017.08.008 [doi]
PST - ppublish
SO  - Zhonghua Fu Chan Ke Za Zhi. 2017 Aug 25;52(8):545-550. doi: 
      10.3760/cma.j.issn.0529-567X.2017.08.008.