Search Thermo Fisher Scientific
Search Thermo Fisher Scientific
Counted by _____________________________ Hepatocyte lot no. _________________________
Date _____________________ Project ID ___________________
Trypan blue counting results
Quadrant | Live cells | + | Dead cells | = | Total cells |
---|---|---|---|---|---|
Quadrant 1 | + | = | |||
Quadrant 2 | + | = | |||
Quadrant 3 | + | = | |||
Quadrant 4 | + | = | |||
Total | + | = |
Viability calculation
Live cells / Total cells = Viability %
/ =
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Total yield calculation
Total cells / Quadrants counted x Dilution factor x 10,000 x Current volume (mL) = Total cell yield
/ x 2 x 10,000 x = x 106 cells
Cell morphology notes
Clear cytoplasm
Rounded shape
Clear/crisp membranes
Minimal debris
Swollen organelles
Membrane blebbing
Lipid droplets
Prevalent debris
Comments: _________________________________________________________________________
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Seeding density calculation
Total cell yield / Desired density* = Total volume needed
x 106 cells / x x 106 cells/mL = mL
Total volume needed -- Current total volume = Volume to add to cell stock
mL -- mL= mL
* Refer to supplier recommendations. Desired density for human plated hepatocytes is typically 0.75 x 106 cells/mL, 0.8 x 106 cells/mL, or 0.9 x 106 cells/mL for a 24-well plate. Densities for suspension use or other species will vary.
Overlay calculation
Total volume of 4°C medium needed* x Final gel concentration / initial protein concentration† = Volume of gel stock to add to 4°C medium
mL x 0.35 mg/mL / mL = mL
* Medium needs to be kept at 4°C on ice. We recommend Williams Medium E (Life Technologies Cat. No. CM6000) combined with a serum-free Maintenance Supplement Pack (Life Technologies Cat. No. CM4000).
† Refer to supplier’s specification sheet. We recommend Geltrex™ Reduced Growth Factor Basement Membrane Matrix (Invitrogen Cat. No. 12760-021) at a concentration of 0.35 mg/mL.
Need assistance? For technical support on hepatocyte products, visit us at www.lifetechnologies.com/hepatocytes,
email us at hepaticproducts@lifetech.com, or contact us by phone at: +1 866 952 3559 (US toll-free); +1 919 237 4500 (toll); +44 (0)141 814 5900 (Europe).
Advance preparation for cryopreserved hepatocytes
Note: Use universal safety precautions and the appropriate biosafety cabinet when handling primary hepatocytes.
Preparation of the cell counting vial
For Research Use Only. Not for use in diagnostic procedures.