By nature iPSCs dislike being "single cells" as it will induce their differentiation without the paracrine factors of adjacent cells, and if you plan on expanding them from single cells they will form colonies anyway.

Can you further specify exactly what you plan on doing with the iPSCs?

Most differentiation protocols will want a dense monolayer culture at minimum for differentiation, others will insist you differentiate into an embryoid body with specific patterning factors to generate progenitors, which you can then further derive specific cell types from.

You can use Accutase to make Single Cells, but must add rock inhibitor 5-10 uM in the medium and after 24 hours of passage change medium without Rock,. When you are working with single cells must use Rock inhibitor or Thiazovivin do increase the viability.

You can use ESC homemade medium if are working on MEF (FEEDER), but after 1 week must make passage or use ESC medium conditionated.

We use E8 or mTeSR1 medium with geltrex or matrigel matrix.

thank you James and Fabiano, both of you.

In fact I wanna transduce iPSCs with lentivirus, and I wanna see if the single cells would be transduced better than colonies.

But even, I'm not sure  if single cells have iPSCs nature or not.

Just one ESC or iPCS single cell will grow and form a new colony like others (same that fragment passage). After it grow up you cant know if come from single cell or fragment.

See if helps:

https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3788326/

Dear Maliheh

In our experience, culturing cells as uniform spheroids is mandatory for proper gene regulation. Hanging drops are the gold standard, but very hard to do im terms of work force needed. If you are interested, we have developed a product for stem cell transplantation which is very useful in lab scale format, too. You are 60x faster than with the fastest multichannel repeater and hanging drops. www.kugelmeiers.com.

If you have any questions, please do not hesitate to contact me.

Regards, Patrick