PMID- 33732326
OWN - NLM
STAT- PubMed-not-MEDLINE
LR  - 20210320
IS  - 1792-0981 (Print)
IS  - 1792-1015 (Electronic)
IS  - 1792-0981 (Linking)
VI  - 21
IP  - 4
DP  - 2021 Apr
TI  - Differentiation of HL-60 cells in serum-free hematopoietic cell media enhances 
      the production of neutrophil extracellular traps.
PG  - 353
LID - 10.3892/etm.2021.9784 [doi]
LID - 353
AB  - Neutrophil extracellular traps (NETs) are web-like structures made of chromatin 
      and have been identified to have a role in the host's immune defense. 
      Differentiated human promyelocytic leukemia HL-60 cells (dHL-60) have been used 
      to study the mechanisms of NETs formation, as neutrophils have a short lifespan 
      that limits their use. However, dHL-60 cells are inefficient at generating NETs 
      and therefore are not ideal replacements for neutrophils in studying of NET 
      formation. In the present study, the optimal cell culture conditions and 
      differentiation time that result in the most effective release of NETs from 
      dHL-60 cells upon stimulation were determined. HL-60 cells were cultured in serum 
      (FBS) or serum-free (X-VIVO) medium and differentiated using all-trans retinoic 
      acid (ATRA) or dimethyl sulfoxide (DMSO). dHL-60 cells were stimulated with 
      phorbol 12-myristate 13-acetate (PMA) or Ca(2+) ionophore (CI). Cell 
      differentiation and apoptosis, as well as the formation of reactive oxygen 
      species (ROS) and citrullinated histone H3 (citH3) were analyzed using flow 
      cytometry. NETs were visualized using fluorescence microscopy and NET 
      quantification was performed using PicoGreen. Induction of HL-60 cells for five 
      days produced the best results in terms of differentiation markers and cell 
      viability. Both ATRA- and DMSO-induced dHL-60 cells were able to release NETs 
      upon PMA and CI stimulation; dHL-60 cells in serum-free medium produced more NETs 
      than those in serum-containing medium. DMSO-dHL-60 (X-VIVO) cells were most 
      efficient at producing NETs and ROS upon stimulation with PMA, while ATRA-dHL-60 
      (X-VIVO) cells were most efficient at producing NETs and citH3 upon stimulation 
      with CI. It was concluded that DMSO-dHL-60 (X-VIVO) may be a model for the study 
      of ROS-high NETosis and ATRA-dHL-60 (X-VIVO) may be suitable for ROS-low NETosis.
CI  - Copyright: (c) Guo et al.
FAU - Guo, Yun
AU  - Guo Y
AD  - Department of Respiratory Medicine, The Affiliated Wuxi Children's Hospital of 
      Nanjing Medical University, Wuxi, Jiangsu 214000, P.R. China.
FAU - Gao, Fei
AU  - Gao F
AD  - Department of Intensive Care Unit, Wuxi People's Hospital Affiliated to Nanjing 
      Medical University, Wuxi, Jiangsu 214000, P.R. China.
FAU - Wang, Qian
AU  - Wang Q
AD  - Department of Respiratory Medicine, The Affiliated Wuxi Children's Hospital of 
      Nanjing Medical University, Wuxi, Jiangsu 214000, P.R. China.
FAU - Wang, Kang
AU  - Wang K
AD  - Department of Laboratory, The Affiliated Wuxi Children's Hospital of Nanjing 
      Medical University, Wuxi, Jiangsu 214000, P.R. China.
FAU - Pan, Shanshan
AU  - Pan S
AD  - Department of Respiratory Medicine, The Affiliated Wuxi Children's Hospital of 
      Nanjing Medical University, Wuxi, Jiangsu 214000, P.R. China.
FAU - Pan, Zhenzhen
AU  - Pan Z
AD  - Department of Respiratory Medicine, The Affiliated Wuxi Children's Hospital of 
      Nanjing Medical University, Wuxi, Jiangsu 214000, P.R. China.
FAU - Xu, Shiyao
AU  - Xu S
AD  - Department of Respiratory Medicine, The Affiliated Wuxi Children's Hospital of 
      Nanjing Medical University, Wuxi, Jiangsu 214000, P.R. China.
FAU - Li, Ling
AU  - Li L
AD  - Department of Respiratory Medicine, The Affiliated Wuxi Children's Hospital of 
      Nanjing Medical University, Wuxi, Jiangsu 214000, P.R. China.
FAU - Zhao, Deyu
AU  - Zhao D
AD  - Department of Respiratory Medicine, Children's Hospital of Nanjing Medical 
      University, Nanjing, Jiangsu 210000, P.R. China.
LA  - eng
PT  - Journal Article
DEP - 20210211
PL  - Greece
TA  - Exp Ther Med
JT  - Experimental and therapeutic medicine
JID - 101531947
PMC - PMC7903455
OTO - NOTNLM
OT  - HL-60
OT  - all-trans retinoic acid
OT  - cell differentiation
OT  - dimethyl sulfoxide
OT  - histone citrullination
OT  - neutrophil
OT  - neutrophil extracellular traps
OT  - reactive oxygen species
OT  - serum-free medium
COIS- The authors declare that they have no competing interests.
EDAT- 2021/03/19 06:00
MHDA- 2021/03/19 06:01
PMCR- 2021/02/11
CRDT- 2021/03/18 06:48
PHST- 2020/04/17 00:00 [received]
PHST- 2020/10/28 00:00 [accepted]
PHST- 2021/03/18 06:48 [entrez]
PHST- 2021/03/19 06:00 [pubmed]
PHST- 2021/03/19 06:01 [medline]
PHST- 2021/02/11 00:00 [pmc-release]
AID - ETM-0-0-09784 [pii]
AID - 10.3892/etm.2021.9784 [doi]
PST - ppublish
SO  - Exp Ther Med. 2021 Apr;21(4):353. doi: 10.3892/etm.2021.9784. Epub 2021 Feb 11.