Hi All,

We have decided to isolate endothelial cells from our mouse model and I am in the process of learning and establishing a protocol so i have a few questions:

  • Has anyone used dynabeads with BD's IMAG magnet? we have one in the lab used for other cell separation projects and i was hoping to use it instead of buying a new very expensive magnet.
  • Has anyone tried to propagate and freeze aliquotes of cells for future experiments as is common with HUVECs? 
  • Since typsin damages CD31 (PECAM-1) has anyone tried replacing it with accutase?
  • When growing the cells after antibody selection and separation, should I heat-shock the FBS? It seems the reasonable thing to do yet it is not mentioned in any protocol I've read (and I read many...)
  • Other than my questions, I will really appreciate and be greatful for any other tips and considerations you can offer.

    Maayan

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