Kelly

Frequently Asked Questions

• What are Kelly cells?

  Kelly cell line were originally isolated from human neuroblastoma and derived from a tumor biopsy.

• What are the applications of Kelly cell line?

  Kelly cells can grow in a monolayer, making them suitable for a wide range of experimental applications. Due to its ability to differentiate into neuron-like cells under specific conditions, Kelly cell line is an identical model for studying the molecular mechanisms of neuroblastoma and potential drug testing fot therapeutic.
  According to the references, glycated Kelly neuroblastoma cells had a much higher potential for migration and invasion compared with non-glycated cells, scientists use Kelly line to study the role of polysialic acid (PSA) in controlling tumor cell growth and differentiation. The Kelly cell line is also a model for studying the role of protein tyrosine phosphatase (SHP1) on Ret oncogene activity inhibition.

• How do I passage Kelly cells?

  Procedure
  Preparation:
  Warm the RPMI1640 medium, PBS, and trypsin-EDTA solution to 37°C in a water bath.
  Ensure all necessary equipment and reagents are sterile and ready for use.

  Observe Cells:
  Place the flask containing ells under an inverted microscope.
  Check for cell confluence (typically, cells should be 70-80% confluent before passaging).

  Remove Old Medium:
  In a biosafety cabinet, aspirate the old medium from the flask carefully without disturbing the cell monolayer.

  Rinse Cells:
  Add 5-10 mL of PBS to the flask to rinse off any residual medium and serum that may inhibit trypsin activity.
  Gently rock the flask to ensure the cells are thoroughly rinsed, then aspirate the PBS.

  Add Trypsin-EDTA:
  Add enough 0.05% trypsin-EDTA solution to cover the cell monolayer (typically 1-2 mL for a T-75 flask).
  Gently tilt the flask to ensure the entire surface is covered with trypsin-EDTA.

  Incubate:
  Place the flask in the incubator for 3-5 minutes, checking the cells under the microscope periodically. The cells should round up and detach from the surface.

  Neutralize Trypsin:
  Once the cells are detached, add 5-10 mL of complete RPMI1640 medium (with FBS) to the flask to neutralize the trypsin.

  Collect Cells:
  Gently pipette the cell suspension up and down to ensure all cells are detached and in suspension.
  Transfer the cell suspension to a 15 mL conical tube.

  Centrifuge:
  Centrifuge the cell suspension at 200 x g for 5 minutes to pellet the cells.

  Resuspend Cells:
  Carefully aspirate the supernatant without disturbing the cell pellet.
  Resuspend the cells in an appropriate volume of fresh complete medium.

  Count Cells (Optional):
  If needed, take a small aliquot of the cell suspension to count cells using a hemocytometer or an automated cell counter.

  Seed New Flasks:
  Seed the cells into new culture flasks or plates at the desired density ( it is recommended to seed at 5×1,000-2×10,000 cells/cm²). For example, if splitting at a 1:5 ratio, add 1 part of the cell suspension to 4 parts of fresh medium in a new flask.

  Incubate:
  Place the newly seeded flasks or plates back into the incubator set at 37°C without CO2 (or use non-vented flask and keep the lid closed).

  Monitoring:
  Observe the cells under the microscope after 24 hours to ensure they have adhered and are spreading out. Change the medium every 2-3 days or as needed.