Improve Research Reproducibility A Bio-protocol resource
bpdetail_icon_lock
Preprint

Culture of MutuDC cell line

KT Kirsteen M Tullett
ML Mireille H Lahoud

Last updated date: Jun 5, 2025 Views: 349 Forks: 0

original An abbreviated version of this protocol was published in eLife in Dec, 2020
RNF41 regulates the damage recognition receptor Clec9A and antigen cross-presentation in mouse dendritic cells
download pdf Download PDF
ask Ask a question
cite How to cite
nfavorite Favorite

Culture of MutuDC (mouse cDC1 line)

 

Cell line: MutuDC 1940

 

Culture Media:

500ml IMDM (Gibco, cat no:12440053)

50ml DC grade Heat Inactivated-FCS (Please note: our FCS is batch tested for DC culture) (In vitro technologies)

5ml Pen/Strep (Sigma, cat no: P0781)

0.25ml 2-ME (Gibco, cat no: 21985023, stock at 55mM, 100x)

Adjust Osmolarity to 308 (± 2) mOsmo/kg using 1 M NaCl 

 

 

Buffers:

Dulbecco PBS (DPBS) (Gibco, cat no 14190250). Adjust osmolarity to 308 mOsmo/kg using 1M NaCl.

 

5mM EDTA in DPBS (308 mOsm/kg)

 

MutuDC culture protocol

 

  1. MutuDC are only semi-adherent.
  2. Collect cell culture supernatant from cell culture flask/plate to a tube.
  3. To dislodge remaining adherent cells from cell culture flask/plate, add 5mM EDTA, incubate for 1-2min at 37C.
  4. Tap flask/use a sterile pipette to detach cells.
  5. Use collected Mutu DC culture supernatant (containing MutuDC from step 2) to rinse flask/plate and pool together.
  6. Spin cells down at 700 x g for 7min, 4C.
  7. Discard supernatant.
  8. Resuspend cells with warmed culture media.
  9. Passage at 1:3 to 1:5 dilution, every 2-3 days. The cells appear healthiest when grown up to 70% confluence as a monolayer. Please do not allow them to overgrow / grow on top of each other.
Copyright: © 2025 The Author(s); This is an open-access article under the CC BY license (https://creativecommons.org/licenses/by/4.0/).
How to cite：
Readers should cite both the Bio-protocol preprint and the original research article where this protocol was used:
  1. Tullett, K and Lahoud, M(2025). Culture of MutuDC cell line. Bio-protocol Preprint. bio-protocol.org/prep2825.
  2. Tullett, K. M., Tan, P. S., Park, H., Schittenhelm, R. B., Michael, N., Li, R., Policheni, A. N., Gruber, E., Huang, C., Fulcher, A. J., Danne, J. C., Czabotar, P. E., Wakim, L. M., Mintern, J. D., Ramm, G., Radford, K. J., Caminschi, I., O'Keeffe, M., Villadangos, J. A., Wright, M. D., Blewitt, M. E., Heath, W. R., Shortman, K., Purcell, A. W., Nicola, N. A., Zhang, J. and Lahoud, M. H.(2020). RNF41 regulates the damage recognition receptor Clec9A and antigen cross-presentation in mouse dendritic cells. eLife. DOI: 10.7554/eLife.63452

Do you have any questions about this protocol?

Post your question to gather feedback from the community. We will also invite the authors of this article to respond.

post Post a Question
0 Q&A
This protocol preprint was submitted via the Bio-protocol Exchange "Submit a Preprint" track.