CoolCell used for GMP-regulated Isolation of Amniotic Stem Cells

Sterile equipment is prepared for the hAEC isolation. Human amnion-derived cells are valued for their plasticity and exceptional safety profile. Image credit: Gramignoli, et al., 2016.

An independent study at the Karolinska Institute in Sweden[1] is using BioCision’s CoolCell® freezing container to aid their research on the isolation and characterization of human amniotic epithelial cells.

The human placenta is a fascinating structure, engineered by evolution to provide both a crucial connection point and an immunological barrier between the developing fetus and the mother. The amnion, an epithelial cell–lined cavity within the placenta, is a rich and generally non-controversial source of progenitor cells. Human amnion epithelial cells (hAECs), for example, can self-renew and differentiate into many kinds of mature cells. They are also non-tumorigenic, and display immunosuppressive properties that make them an attractive candidate for use in regenerative medicine.

We have previously discussed the need for safe and effective methods for cryopreserving cells derived from the human amnion; including the importance of integrating good manufacturing practices and quality control. Because current regulatory guidelines don’t clearly define parameters for isolation and banking of hAECs, this current study from the Karolinska Institute delineates a detailed protocol suitable to clinical applications.

In accordance with GMP recommendations for clinical grade cells, all reagents used in the procedure are animal-free in origin, to avoid the risk of contamination or infectious pathogens. The authors also recommend only using placental tissue derived from delivery by caesarean sections, since this guarantees the tissue was collected under sterile conditions. Following dissection of the amnion away from the placenta, hAECs are released from the tissue by enzymatic digestion, after which cell recovery and viability assays are performed. The cells are then ready for cryopreservation and banking.

Banking clinical grade cells is what allows for the widespread application of human cells in preclinical evaluation and therapeutic treatments. For this reason, the authors believe cryopreservation steps need to be clearly defined and always follow GMP guidelines. Freezing cells by slow cooling is currently considered the best method of achieving high viability and post-thaw recovery rates. The authors use BioCision’s CoolCell for this step, because, as they note, “The freezing container should be alcohol-free and able to provide a freezing rate of −1°C/min when transferred to a −80°C freezer.” In this protocol, cells remain in the −80°C freezer for 90 min. before being transferred to a liquid nitrogen container for long-term storage.

When the hAECs are finally thawed, GMP regulations require for them to be characterized in terms of viability, identity, and function before they can be used. The authors are confident that their protocol will provide a cell product that is useful for both basic research, and therapeutic use in the clinic.

Reference:

[1] Gramignoli R., et al. Isolation of Human Amnion Epithelial Cells According to Current Good Manufacturing Procedures. Current Protocols in Stem Cell Biology. 37:1E.10.1-1E.10.13. May 2016.