PMID- 33836009
OWN - NLM
STAT- MEDLINE
DCOM- 20210915
LR  - 20210915
IS  - 1932-6203 (Electronic)
IS  - 1932-6203 (Linking)
VI  - 16
IP  - 4
DP  - 2021
TI  - Generation and utilization of a HEK-293T murine GM-CSF expressing cell line.
PG  - e0249117
LID - 10.1371/journal.pone.0249117 [doi]
LID - e0249117
AB  - Macrophages and dendritic cells (DCs) are innate immune cells that play a key 
      role in defense against pathogens. In vitro cultures of bone marrow-derived 
      macrophages (BMDMs) and dendritic cells (BMDCs) are well-established and valuable 
      methods for immunological studies. Typically, commercially available recombinant 
      GM-CSF is utilized to generate BMDCs and is also used to culture alveolar 
      macrophages. We have generated a new HEK-293T cell line expressing murine GM-CSF 
      that secretes high levels of GM-CSF (~180 ng/ml) into complete media as an 
      alternative to commercial GM-CSF. Differentiation of dendritic cells and 
      expression of various markers were kinetically assessed using the GM-CSF HEK293T 
      cell line, termed supGM-CSF and compared directly to purified commercial GMCSF. 
      After 7-9 days of cell culture the supGM-CSF yielded twice as many viable cells 
      compared to the commercial purified GM-CSF. In addition to differentiating BMDCs, 
      the supGM-CSF can be utilized to culture functionally active alveolar 
      macrophages. Collectively, our results show that supernatant from our GM-CSF 
      HEK293T cell line supports the differentiation of mouse BMDCs or alveolar 
      macrophage culturing, providing an economical alternative to purified GM-CSF.
FAU - Robinson, Elektra Kantzari
AU  - Robinson EK
AUID- ORCID: 0000-0003-1427-3897
AD  - Department of Molecular, Cell and Developmental Biology, University of California 
      Santa Cruz, Santa Cruz, CA, United States of America.
FAU - Covarrubias, Sergio
AU  - Covarrubias S
AUID- ORCID: 0000-0002-7197-9952
AD  - Department of Molecular, Cell and Developmental Biology, University of California 
      Santa Cruz, Santa Cruz, CA, United States of America.
FAU - Zhou, Simon
AU  - Zhou S
AUID- ORCID: 0000-0001-8695-7943
AD  - Department of Molecular, Cell and Developmental Biology, University of California 
      Santa Cruz, Santa Cruz, CA, United States of America.
FAU - Carpenter, Susan
AU  - Carpenter S
AD  - Department of Molecular, Cell and Developmental Biology, University of California 
      Santa Cruz, Santa Cruz, CA, United States of America.
LA  - eng
GR  - R21 AR070973/AR/NIAMS NIH HHS/United States
PT  - Journal Article
PT  - Research Support, N.I.H., Extramural
DEP - 20210409
PL  - United States
TA  - PLoS One
JT  - PloS one
JID - 101285081
RN  - 0 (Culture Media, Conditioned)
RN  - 83869-56-1 (Granulocyte-Macrophage Colony-Stimulating Factor)
SB  - IM
MH  - Animals
MH  - Cell Culture Techniques/methods
MH  - Cell Differentiation
MH  - Culture Media, Conditioned/metabolism/pharmacology
MH  - Dendritic Cells/cytology/drug effects
MH  - *Exocytosis
MH  - Granulocyte-Macrophage Colony-Stimulating Factor/genetics/*metabolism
MH  - HEK293 Cells
MH  - Humans
MH  - Mice
MH  - Transgenes
PMC - PMC8034741
COIS- The authors have declared that no competing interests exist.
EDAT- 2021/04/10 06:00
MHDA- 2021/09/16 06:00
PMCR- 2021/04/09
CRDT- 2021/04/09 17:16
PHST- 2020/06/18 00:00 [received]
PHST- 2021/03/11 00:00 [accepted]
PHST- 2021/04/09 17:16 [entrez]
PHST- 2021/04/10 06:00 [pubmed]
PHST- 2021/09/16 06:00 [medline]
PHST- 2021/04/09 00:00 [pmc-release]
AID - PONE-D-20-18789 [pii]
AID - 10.1371/journal.pone.0249117 [doi]
PST - epublish
SO  - PLoS One. 2021 Apr 9;16(4):e0249117. doi: 10.1371/journal.pone.0249117. 
      eCollection 2021.