Practical and Safe Method of Long-Term Cryopreservation for Clinical Application of Human Adipose-Derived Mesenchymal
1Department of Transfusion Medicine, Aichi Medical University, Nagakute, Aichi 480-1195, Japan. sgao0303@aichi-med-u.ac.jp.
Background:
Adipose-derived mesenchymal stem cells (ADSCs) have emerged as a promising modality for cellular therapy. However, techniques of ADSC cryopreservation, which can facilitate their clinical application, haven't been established yet.
Objective:
To determine optimal conditions for ADSC cryopreservation.
Materials And Methods:
We used three cryoprotectants [serum containing 10% dimethyl sulfoxide; CP-1TM (5% dimethyl sulfoxide, serum-free); Stem-CellBankerTM (dimethyl sulfoxide and serum-free)], two storage temperatures (-80°C, -150°C) and two cell densities (1 × 106, 7 × 106 cells/mL). Storage was up to 18 months using cryovials. We didn't use a rate-controlled freezer or liquid nitrogen storage.
Results:
We found that CP-1TM was a suitable cryoprotectant. Storage at -150°C and higher cell density (7×106 cells/mL) kept the best viability of ADSCs, but storage at -80°C and a lower cell density (1×106 cells/mL) is acceptable for up to 9 months. We also confirmed large quantities of ADSCs, stored with CP-1 in a cryobag, were still viable after -150°C cryopreservation for 24 months.
Conclusion:
We have developed a safe, cost-effective way to cryopreserve ADSCs that could be used in the clinical setting.
