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Dear author,I am a PHD student in biomolecular medicine at University...

Giacomo Foschi Mar 15, 2019

Protocol: Culture of Megakaryocytes from Human Peripheral Blood Mononuclear Cells

Dear author,

I am a PHD student in biomolecular medicine at University of Verona and II would like to ask you a question about cell culturing.
When you washed/refreshed the cells, did you put in new wells the same density (1-2 x 106 cells/ml) used at The beginning of the ce Read more more

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Petros Papadopoulos Author Answered Mar 15, 2019

Hematology, Hospital Clinico San Carlos , Madrid

Yes, that’s correct.

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Giacomo Foschi Answered Mar 15, 2019

University of Verona

Dear Petros,

Thank you very much for your explanation.
One more question:
you suggested to manipulate cells as less as possible after 3/4 days. Therefore, in order to start phase 2 or phase 3, It is better to add phase 2/3 medium to old medium without pelleting cells, isn't it?

Thank you.
Best, Giacomo

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Petros Papadopoulos Author Answered Mar 15, 2019

Hematology, Hospital Clinico San Carlos , Madrid

Dear Giacomo, in principle you should keep the same conc, yes. You can either replate the cells OR just refresh them on the same plate. Replating is better in the first 3-4 days of the culture, later on its better to keep the culture with less manipulation possible, just add medium.Best.

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