Jun 25, 2024

Public workspaceHuman Embryonic Kidney Cells (HEK-293)

DOI
dx.doi.org/10.17504/protocols.io.x54v92741l3e/v1
  • Md Razaul Karim1
  • 1University of Minnesota
  • Md Razaul Karim: Lee Lab
Jane Balster
Open access
DOI: dx.doi.org/10.17504/protocols.io.x54v92741l3e/v1
Protocol Citation: Md Razaul Karim 2024. Human Embryonic Kidney Cells (HEK-293). protocols.io https://dx.doi.org/10.17504/protocols.io.x54v92741l3e/v1
License: This is an open access protocol distributed under the terms of the Creative Commons Attribution License,  which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited
Protocol status: Working
We use this protocol and it's working
Created: June 07, 2024
Last Modified: June 25, 2024
Protocol Integer ID: 102394
Keywords: Human Embryonic Kidney Cells (HEK-293), DMEM, Drug treat, Harvesting
Funders Acknowledgement:
ASAP
Grant ID: 000592
Abstract
This protocol details the Human Embryonic Kidney Cells (HEK-293) culture.
Materials
Media

  • DMEM full media containing (DMEM/10% FBS/1% Pen-Strep).

Materials

  • 12 Well plate (1 ml/well)
  • 6 Well plate/35 mm dish (9.5 cm2-2 ml/well)
  • 60 mm dish (21 cm2-3.5 ml/dish)
  • 100 mm dish (56 cm2-10 ml/dish)
  • 250 ml Flask for sub-culture/maintenance (10 ml) [Tissue culture flask-Greiner bio-one- Cat.No.-658 170]ReagentCELL CULTURE FLASK, 250 ML, 75 CM², PS, RED STANDARD SCREW CAP, CLEAR, CELLSTAR® TC, STERILE, 5 PCS.greiner bio-oneCatalog #658170

Day-01 (Mon)
Day-01 (Mon)
36m
Plating with DMEM full media .
Warm DMEM full media, PBS, and Trypsin in the Temperature37 °C bead bath for Duration00:30:00 . Clean the working area by using 70% ethanol.
30m
Temperature
Sup out old media without touching cells.
Wash by adding Amount5 mL PBS slowly, rinse, and rock back and forth.
Pipetting
Wash
Add Amount2 mL -Amount3 mL trypsin (0.25%); keep in incubator for Duration00:03:00 .
3m
Incubation
Pipetting
Check under microscope if cells are detached, add Amount5 mL media and transfer to a tube.
Pipetting
Imaging
Spin Shaker300 x g, 00:03:00 for Duration00:03:00 .
3m
Mix
Sup out and add Amount10 mL fresh media & re-suspend cells gently and carefully.
Pipetting
Count cells density and split accordingly. 15,000 cells/ml for maintenance.

  • (i) Usually 1.0-1.5x10^4/ml cells for Biochem, and
  • (ii) 0.5x10^4/ml cells for IF.
Day-02 (Tue)
Day-02 (Tue)
Rest.
Day-03 (Wed)
Day-03 (Wed)
Replace with DMEM full media/Drug treat.
Day-04 (Thu)
Day-04 (Thu)
Drug treat if necessary /Harvesting.
Day-05 (Fri)
Day-05 (Fri)
Drug treat if necessary /Harvesting.