====== Original protocol ======
===== from Michael Heide lab (Göttingen) =====
iPSC culture (mTeSR1) 

Protocol iPSC culture for chimpanzee iPSCs and ZNF492-WT and KO cells

Coating of plates

  * thaw Geltrex on ice
  * dilute Geltrex in cold DMEM/F12 1:100 (for 1 well of a 6-well plate 1 ml diluted Geltrex is needed)
  * incubate for 1 h. at RT

Thawing of iPS cells

  * coat plates with Geltrex
  * prewarm mTeSR1 medium
  * prepare 4 ml mTeSR1 in 15 ml falcon tube
  * thaw cells at 37°C
  * move cells to prepared falcon tube
  * centrifuge for 5 min
  * remove Geltrex from coated plates and add 1 ml mTeSR1 per well
  * remove supernatant
  * resuspend cells in 1 ml mTeSR1 using a 5 ml serological pipette
  * move cells to coated plate
  * incubate at 37°C and 5% CO2
  * daily medium changes

Medium Change

  * prewarm mTeSR
  * remove old medium
  * add 2 ml mTeSR1
  * incubate at 37°C and 5% CO2

Passaging of iPS cells

  * coat plates with Geltrex
  * prewarm mTeSR1 medium
  * wash cells with PBS
  * add 1 ml ReLeSR (at RT)
  * remove ReLeSR
  * incubate in incubator for 3 min.
  * add 1 ml mTeSR1 and tap for 30 sec.
  * move cells to a falcon tube and add 5 ml mTeSR1 for 1:6 split
  * remove Geltrex from the coated plates and add 1 ml mTeSR1 per well
  * move 1 ml cell suspension to each well of the plate
  * incubate at 37°C and 5% CO2
  * daily medium changes

Freezing of iPS cells

  * thaw mFreSR1
  * wash cells with PBS
  * add 1 ml ReLeSR (at RT)
  * remove ReLeSR
  * incubate in incubator for 3 min.
  * add 1 ml mFreSR1 and tap for 30 sec.
  * move cell suspension to a cryotube
  * freeze in a Mr. Frosty at -80°C