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Cell aging
Platingdensityistoolow
Somestemcellscansecretefactorsto
supportcellgrowth.Therefore,acertain
degreeofplatingdensitymustbe
maintained;otherwise,itwillleadtocell
Troubleshooting
The table below lists some potential problems and solutions for culturing ADSCs.
Problem
Low cell recovery
rate
Slow cell growth
Cause
Solution
Thestorageconditiondoes
notmeettherequirements
Purchaseareplacementandstoreinliquid
nitrogenforlongͲtermpreservation.
Thawingofthecellstakestoo
long
Thawcellsfornomorethan3minutes.
Cellsareincompletely
recoveredafterthawing
Afteraspiratingoffmedium,washthe
tubewithculturemediumtwiceand
transferallofthecellstothedish.
Cellsarehandledroughly
Careshouldbetakentoavoidintroducing
bubblesduringpipetting.Alsoavoid
vortexingandhighͲspeedcentrifugation
MediumisnotpreͲwarmed
Warmmediumto37°Cbeforerecovery.
Mycoplasmacontamination
Discardthecellsinquestionanddisinfect
thelaboratoryenvironmentbefore
recoveringthenextbatchofcells.
Overdigestion
WashthecellswithPBS2Ͳ3timesto
removeserumpriortotrypsinization(serum
willinhibitthefunctionoftrypsin).
Controlthe digestiontime.
Cell aging
Platingdensityistoolow
Increasetheplatingdensity.
Inappropriateserumand
medium
UseCyagentailorͲmadeculturemedia. If
otherserumandmediaproductsareused,
pleaseperformvalidationtoensure
compatibility.
Deadcellsarenotremoved
promptly
Changethemediumthenextdayafter
recoverytoensureremovalofalldead
cells.
CellContamination
Discardthecellsinquestionanddisinfect
thelaboratoryenvironmentbefore
recoveringthenextbatchofcells.
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