PMID- 35311827
OWN - NLM
STAT- MEDLINE
DCOM- 20220407
LR  - 20220407
IS  - 1940-087X (Electronic)
IS  - 1940-087X (Linking)
IP  - 181
DP  - 2022 Mar 7
TI  - Study of Dendritic Cell Development by Short Hairpin RNA-Mediated Gene Knockdown 
      in a Hematopoietic Stem and Progenitor Cell Line In vitro.
LID - 10.3791/62730 [doi]
AB  - Dendritic cells (DCs) are important antigen-presenting cells that connect innate 
      and adaptive immune responses. DCs are heterogeneous and can be divided into 
      conventional DCs (cDCs) and plasmacytoid DCs (pDCs). cDCs specializes in 
      presenting antigens to and activate naive T cells. On the other hand, pDCs can 
      produce large quantities of type I interferons (IFN-I) during viral infection. 
      The specification of DCs occurs at an early stage of DC progenitors in the bone 
      marrow (BM) and is defined by a network of transcription factors (TFs). For 
      example, cDCs highly express ID2, while pDCs highly express E2-2. Since more and 
      more subsets of DCs are being identified, there is a growing interest in 
      understanding specific TFs controlling DC development. Here, we establish a 
      method to screen TFs critical for DCs differentiation in vitro by delivering 
      lentivirus carrying short hairpin RNA (shRNA) into an immortalized hematopoietic 
      stem and progenitor cell (iHSPCs) line. After the selection and in vitro 
      differentiation, cDC and pDC potential of the stable knockdown cell lines are 
      analyzed by flow cytometry. This approach provides a platform to identify genes 
      potentially governing DC fates from progenitors in vitro.
FAU - Hsiao, Yu-Ling
AU  - Hsiao YL
AD  - Graduate Institute of Immunology, National Taiwan University College of Medicine.
FAU - Hacker, Hans
AU  - Hacker H
AD  - Department of Pathology, Division of Microbiology and Immunology, University of 
      Utah.
FAU - Lee, Chien-Kuo
AU  - Lee CK
AD  - Graduate Institute of Immunology, National Taiwan University College of Medicine; 
      leeck@ntu.edu.tw.
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PT  - Video-Audio Media
DEP - 20220307
PL  - United States
TA  - J Vis Exp
JT  - Journal of visualized experiments : JoVE
JID - 101313252
RN  - 0 (RNA, Small Interfering)
SB  - IM
MH  - Cell Differentiation
MH  - Cell Line
MH  - *Dendritic Cells
MH  - Gene Knockdown Techniques
MH  - *Hematopoietic Stem Cells
MH  - RNA, Small Interfering/genetics/metabolism
EDAT- 2022/03/22 06:00
MHDA- 2022/04/08 06:00
CRDT- 2022/03/21 12:17
PHST- 2022/03/21 12:17 [entrez]
PHST- 2022/03/22 06:00 [pubmed]
PHST- 2022/04/08 06:00 [medline]
AID - 10.3791/62730 [doi]
PST - epublish
SO  - J Vis Exp. 2022 Mar 7;(181). doi: 10.3791/62730.