1. Home
  2. Protocol
  3. Primary Cell Protocol
  4. Protocol Adipocyte Differentiation for Umbilical Cord Derived MSCs

Protocol Adipocyte Differentiation for Umbilical Cord Derived MSCs

Product Information

Adipocyte Differentiation for Umbilical Cord Derived MSCs

Catalog No. Product Name Product quantity Short-term Storage Long-term Storage Thawing Instructions
ax9003 MSCs (Umbilical Cord 500,000 Liquid Nitrogen Liquid Nitrogen See Protocol
Derived) cells/vial
ax9005 MSC Expansion Medium 500 mL 4°C for 1 -20°C for 6 Thaw at 4°C or
for Adipose Tissue month months RT
Derived & Umbilical Cord
Derived MSCs
ax9019 MSC Adipogenesis 100 mL Store at 4°C N/A N/A
Medium for Bone Marrow for up to 6
Derived and Umbilical weeks
Cord Derived MSCs
Aliquot and
ax0049 Fibronectin Coating Solution 1 mL store at -80for up to 3 oC Thaw at 4 C o Once diluted, use
months immediately

Recommendations:

  • Recommended culture vessel coating: Fibronectin
  • Recommended cell culture medium: Axol MSC Expansion Medium followed by Axol MSC Adipogenesis Medium (see Table)
  • Seeding density for Adipogenesis: 40,000-60,000 viable cells/cm2
  • Recommended centrifugation speed: 200 x g for 5 min

Coating:

  • Coat the required number of 6-well plates with Fibronectin Coating Solution.
  • Dilute the stock Fibronectin Coating Solution 1:50 in sterile water to make 1x working solution e.g. 100 μL in 5 mL.
  • On the day prior to thawing the cells, coat the surface of your culture vessel with the Fibronectin 1x working solution. We recommend coating at a volume of 200 μL per cm2 however, please optimize for your experiments.
  • Incubate the culture vessel overnight at 37°C in a humidified incubator.
  • Remove the coating solution prior to seeding the cells.

Coating the culture vessels with Fibronectin will prevent the cells from detaching during differentiation

Seeding:

  • Expand the MSCs (Umbilical Cord Derived) in MSC Expansion Medium for Adipose
  • Tissue Derived & Umbilical Cord Derived MSCs until the cells are actively proliferating.
  • Passage when the culture reaches: 70-90% confluent
  • Recommended passaging reagent: Trypsin-EDTA
  • Neutralize the trypsin with pre-warmed cell culture medium and centrifuge the cells at 200x g for 5 min.
  • Remove the supernatant and resuspend in 1-2 mL of pre-warmed cell culture medium.
  • Perform a cell count to determine the number of viable cells.
  • Seed cells into the 6-well plate (coated with Fibronectin) at the recommended seeding density (40,000-60,000 viable cells/cm2 ) in  pre-warmed MSC Expansion Medium (2 mL per well for a 6-well plate).
  • Incubate the cells at 37°C, 5% CO2 in a humidified incubator.

Usage Statement:

Our products are intended for research use only and are not to be used for any other purpose, which includes but is not limited to, unauthorized commercial uses, in vitro diagnostic uses, ex vivo or in vivo therapeutic uses or any type of consumption or application to humans.