Efficient aseptic and automatable vitrification of human Embryonic Stem Cells using bio-safe & chemically defined media

We have demonstrated on human pluripotent stem cells that, despite additional constraints (aseptic, defined medium, automatable…), our aseptic (i.e. in closed systems) and chemically defined (i.e. without complex undefined compinent) vitrification method is more efficient than conventional slow freezing. We can also conclude that the cells keep their biological (pluripotency) properties after aseptic vitrification, including a normal karyotype and the capacity to generate teratoma when injected in immunodeficient mice.
Aseptic vitrification of human Pluripotent Stem Cells is now possible in completely defined media. Because straws are directly immersed in LN2, this method does not require any specific and expensive material. To limit cell manipulations, our protocol implies stepwise addition and dilution of cryoprotectants before cooling and after warming, respectively, allowing automation.