Gao S, Ogawa M, Takami A, Takeshita K, Kato H
Department of Transfusion Medicine, Aichi Medical University, Nagakute, Aichi 480-1195, Japan.
Clinical Laboratory, Aichi Medical University, Nagakute, Aichi 480-1195, Japan.
Cryo Letters. 2020 Nov-Dec;41(6):337-343.
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.
To determine optimal conditions for ADSC cryopreservation.
We used three cryoprotectants [serum containing 10% dimethyl sulfoxide; CP-1 (5% dimethyl sulfoxide, serum-free); Stem-CellBanker (dimethyl sulfoxide and serum-free)], two storage temperatures (-80°C, -150°C) and two cell densities (1 × 10, 7 × 10 cells/mL). Storage was up to 18 months using cryovials. We didn't use a rate-controlled freezer or liquid nitrogen storage.
We found that CP-1 was a suitable cryoprotectant. Storage at -150°C and higher cell density (7×10 cells/mL) kept the best viability of ADSCs, but storage at -80°C and a lower cell density (1×10 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.
We have developed a safe, cost-effective way to cryopreserve ADSCs that could be used in the clinical setting.
脂肪来源的间充质干细胞（ADSCs）已成为一种有前景的细胞治疗方式。然而，尚未建立能促进其临床应用的ADSC冷冻保存技术。
确定ADSC冷冻保存的最佳条件。
我们使用了三种冷冻保护剂[含10%二甲基亚砜的血清；CP-1（5%二甲基亚砜，无血清）；干细胞库保存液（二甲基亚砜且无血清）]、两种储存温度（-80°C、-150°C）和两种细胞密度（1×10、7×10个细胞/毫升）。使用冻存管储存长达18个月。我们未使用速率控制冷冻仪或液氮储存。
我们发现CP-1是一种合适的冷冻保护剂。在-150°C储存且细胞密度较高（7×10个细胞/毫升）时，ADSCs的活力保持最佳，但在-80°C且细胞密度较低（1×10个细胞/毫升）下储存长达9个月也是可以接受的。我们还证实，用CP-1保存在冻存袋中的大量ADSCs在-150°C冷冻保存24个月后仍具有活力。
我们已开发出一种安全、经济高效的ADSCs冷冻保存方法，可用于临床。
