PMID- 36738210
OWN - NLM
STAT- MEDLINE
DCOM- 20230207
LR  - 20230207
IS  - 1872-2075 (Electronic)
IS  - 1000-3061 (Linking)
VI  - 39
IP  - 1
DP  - 2023 Jan 25
TI  - [Development of porcine induced pluripotent stem cells with a CD163 reporter 
      system].
PG  - 192-203
LID - 10.13345/j.cjb.220288 [doi]
AB  - As main recipient cells for porcine reproductive and respiratory syndrome virus 
      (PRRSV), porcine alveolar macrophage (PAM) are involved in the progress of 
      several highly pathogenic virus infections. However, due to the fact that the PAM 
      cells can only be obtained from primary tissues, research on PAM-based virus-host 
      interactions remains challenging. The improvement of induced pluripotent stem 
      cells (iPSCs) technology provides a new strategy to develop IPSCs-derived PAM 
      cells. Since the CD163 is a macrophage-specific marker and a validated receptor 
      essential for PRRSV infection, generation of stable porcine induced pluripotent 
      stem cells lines containing CD163 reporter system play important roles in the 
      investigation of IPSCs-PAM transition and PAM-based virus-host interaction. Based 
      on the CRISPR/Cas9- mediated gene editing system, we designed a sgRNA targeting 
      CD163 locus and constructed the corresponding donor vectors. To test whether this 
      reporter system has the expected function, the reporter system was introduced 
      into primary PAM cells to detect the expression of RFP. To validate the low 
      effect on stem cell pluripotency, we generated porcine iPSC lines containing 
      CD163 reporter and assessed the pluripotency through multiple assays such as 
      alkaline phosphatase staining, immunofluorescent staining, and EdU staining. The 
      red-fluorescent protein (RFP) expression was detected in CD163-edited PAM cells, 
      suggesting that our reporter system indeed has the ability to reflect the 
      expression of gene CD163. Compared with wild-type (WT) iPSCs, the CD163 
      reporter-iPSCs display similar pluripotency-associated transcription factors 
      expression. Besides, cells with the reporter system showed consistent cell 
      morphology and proliferation ability as compared to WT iPSCs, indicating that the 
      edited-cells have no effect on stem cell pluripotency. In conclusion, we 
      generated porcine iPSCs that contain a CD163 reporter system. Our results 
      demonstrated that this reporter system was functional and safe. This study 
      provides a platform to investigate the iPS-PAM development and virus-host 
      interaction in PAM cells.
FAU - Yue, Wei
AU  - Yue W
AD  - Shaanxi Stem Cell Engineering and Technology Research Center, College of 
      Veterinary Medicine, Northwest A & F University, Yangling 712100, Shaanxi, China.
FAU - Zhang, Juqing
AU  - Zhang J
AD  - Shaanxi Stem Cell Engineering and Technology Research Center, College of 
      Veterinary Medicine, Northwest A & F University, Yangling 712100, Shaanxi, China.
FAU - Wu, Xiaolong
AU  - Wu X
AD  - Shaanxi Stem Cell Engineering and Technology Research Center, College of 
      Veterinary Medicine, Northwest A & F University, Yangling 712100, Shaanxi, China.
FAU - Yang, Xinchun
AU  - Yang X
AD  - Shaanxi Stem Cell Engineering and Technology Research Center, College of 
      Veterinary Medicine, Northwest A & F University, Yangling 712100, Shaanxi, China.
FAU - Shen, Qiaoyan
AU  - Shen Q
AD  - Shaanxi Stem Cell Engineering and Technology Research Center, College of 
      Veterinary Medicine, Northwest A & F University, Yangling 712100, Shaanxi, China.
FAU - Yu, Shuai
AU  - Yu S
AD  - Shaanxi Stem Cell Engineering and Technology Research Center, College of 
      Veterinary Medicine, Northwest A & F University, Yangling 712100, Shaanxi, China.
FAU - Zhu, Zhenshuo
AU  - Zhu Z
AD  - Shaanxi Stem Cell Engineering and Technology Research Center, College of 
      Veterinary Medicine, Northwest A & F University, Yangling 712100, Shaanxi, China.
FAU - Wang, Chengbao
AU  - Wang C
AD  - Department of Preventive Veterinary Medicine, College of Veterinary Medicine, 
      Northwest A & F University, Yangling 712100, Shaanxi, China.
FAU - Zhang, Shiqiang
AU  - Zhang S
AD  - Shaanxi Stem Cell Engineering and Technology Research Center, College of 
      Veterinary Medicine, Northwest A & F University, Yangling 712100, Shaanxi, China.
FAU - Hua, Jinlian
AU  - Hua J
AD  - Shaanxi Stem Cell Engineering and Technology Research Center, College of 
      Veterinary Medicine, Northwest A & F University, Yangling 712100, Shaanxi, China.
LA  - chi
PT  - English Abstract
PT  - Journal Article
PL  - China
TA  - Sheng Wu Gong Cheng Xue Bao
JT  - Sheng wu gong cheng xue bao = Chinese journal of biotechnology
JID - 9426463
RN  - 0 (CD163 antigen)
RN  - 0 (Receptors, Cell Surface)
RN  - 0 (Antigens, CD)
SB  - IM
MH  - Swine
MH  - Animals
MH  - *Induced Pluripotent Stem Cells/metabolism
MH  - Receptors, Cell Surface/genetics
MH  - Antigens, CD/genetics/metabolism
MH  - *Porcine respiratory and reproductive syndrome virus/genetics
OTO - NOTNLM
OT  - CD163
OT  - CRISPR/Cas9
OT  - improvement of induced pluripotent stem cells (iPSCs)
OT  - porcine
OT  - porcine reproductive and respiratory syndrome virus (PRRSV)
OT  - reporter vector
EDAT- 2023/02/05 06:00
MHDA- 2023/02/08 06:00
CRDT- 2023/02/04 06:12
PHST- 2023/02/04 06:12 [entrez]
PHST- 2023/02/05 06:00 [pubmed]
PHST- 2023/02/08 06:00 [medline]
AID - 10.13345/j.cjb.220288 [doi]
PST - ppublish
SO  - Sheng Wu Gong Cheng Xue Bao. 2023 Jan 25;39(1):192-203. doi: 
      10.13345/j.cjb.220288.