The cross-talk between ROS and p38MAPKα in the Ex Vivo expanded human umbilical cord blood CD133(+) cells.
This study investigated the correlation between and compared the effects of reactive oxygen species (ROS) and p38 mitogen-activated protein kinase α (p38MAPKα) in the ex vivo expanded umbilical cord blood (hUCB) CD133(+) cells. hUCB CD133(+) cells were cultured in the hematopoietic stem cells (HSCs) culture medium with N-acetylcysteine (NAC, an anti-oxidant), p38MAPKα-specific inhibitor (SB203580) or their combination. The levels of ROS and expression of phosphorylated p38MAPKα (p-p38) in CD133(+) cells were flow cytometrically detected. The efficacy of ex vivo expansion was evaluated by the density of CD133(+) cell sub-group colony-forming cells (CFC) and cobblestone area-forming cells (CAFC) assay. Our results showed decreased ROS levels in NAC, SB203580, and their combination treatment groups were almost 37%, 48%, and 85%, respectively. Furthermore, SB203580 abrogated the activation of p38MAPKα more obviously than NAC. Moreover, the CD133(+) cells in SB203580 treatment group had a 21.93±1.36-fold increase, and 14.50±1.19-fold increase in NAC treatment group, but only 10.13±0.57-fold increase in control group. In addition, SB203580 treatment led a higher level increase in the number of CFU and CAFC than NAC did. These findings suggested that, in expanded CD133(+) cells, ROS activates p38MAPKα, which, in turn, induces ROS production, and p38MAPKα might be the most suitable regulator in ROS-p38MAPKα pathway for the promotion of HSCs ex vivo expansion.
Department of Hematology, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, 430022, China.
Jing
J
Zou
Department of Hematology, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, 430022, China. xys387@163.com.
Ping
P
Zou
Department of Hematology, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, 430022, China.
Yi
Y
Lou
Department of Hematology, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, 430022, China.
Yin
Y
Xiao
Department of Hematology, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, 430022, China.
Jie
J
Wang
Department of Hematology, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, 430022, China. liulingbo64@yahoo.com.
Lingbo
L
Liu
D000071916:AC133 Antigen; D000111:Acetylcysteine / Q000494:pharmacology; D015703:Antigens, CD / Q000097:blood*; D002455:Cell Division; D002478:Cells, Cultured; D003470:Culture Media; D005312:Fetal Blood / Q000166:cytology*; D006023:Glycoproteins / Q000097:blood*; D006412:Hematopoietic Stem Cells / Q000166:cytology*; D006801:Humans; D007093:Imidazoles / Q000494:pharmacology; D048308:Mitogen-Activated Protein Kinase 14 / Q000378:metabolism*; D010455:Peptides / Q000097:blood*; D047428:Protein Kinase Inhibitors / Q000494:pharmacology; D011725:Pyridines / Q000494:pharmacology; D017382:Reactive Oxygen Species / Q000378:metabolism*
D000071916:AC133 Antigen; D015703:Antigens, CD; D003470:Culture Media; D006023:Glycoproteins; D007093:Imidazoles; C000606023:PROM1 protein, human; D010455:Peptides; D047428:Protein Kinase Inhibitors; D011725:Pyridines; D017382:Reactive Oxygen Species; D048308:Mitogen-Activated Protein Kinase 14; C093642:SB 203580; D000111:Acetylcysteine