Personal Genome Project 1 (PGP1) iPSCs or, for all EB-derived cardiac OBB protocols, BJFF iPSCs (provided by S. Jain at Washington University) were cultured on Matrigel-coated plates (Corning Life Sciences) and maintained in mTeSR1 medium (STEMCELL Technologies). Once colonies began to merge (60 to 80% confluency), cells were rinsed with phosphate-buffered saline (PBS) without calcium or magnesium and passaged by adding an enzyme-free passaging solution (ReLeSR, STEMCELL Technologies) at 1 ml/10 cm2 of tissue culture area. The ReLeSR was immediately aspirated away, and the flask was incubated for 7 min at 37°C, 5% CO2. To lift off colonies, fresh mTeSR1 (1 ml/10 cm2) was gently added to the flask using a serological pipette and slowly pipetted back up once to break up the colonies. The cells were then added to a freshly prepared Matrigel-coated flask, performing a split between 1:8 and 1:3, and mTeSR1 was added to a total volume of 2 ml/10 cm2 of tissue culture area. Cells were maintained at 37°C/5% CO2 in an incubator.

Note: The content above has been extracted from a research article, so it may not display correctly.

Please log in to submit your questions online.
Your question will be posted on the Bio-101 website. We will send your questions to the authors of this protocol and Bio-protocol community members who are experienced with this method. you will be informed using the email address associated with your Bio-protocol account.

We use cookies on this site to enhance your user experience. By using our website, you are agreeing to allow the storage of cookies on your computer.