Cryopreservation of human mesenchymal stromal cells expressing TRAIL for human anti-cancer therapy

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• 作者：Zhengqiang Yuan¹ ; Sofia Da Silva Lourenco¹ ; Elizabeth K. Sage¹ ; Krishna K. Kolluri¹ ; Mark W. Lowdell² ; Sam M. Janes¹ ; ^{s.janes@ucl.ac.uk" class="auth_mail" title="E-mail the corresponding author}
• 关键词：apoptosis ; chemokine ; cryopreservation ; DMSO ; MSC ; ZENALB 4.5
• 刊名：Cytotherapy
• 出版年：2016
• 出版时间：July 2016
• 年：2016
• 卷：18
• 期：7
• 页码：860-869
• 全文大小：978 K

文摘

Mesenchymal stromal cells (MSCs) are being extensively researched for cell therapy and tissue engineering. We have engineered MSCs to express the pro-apoptotic protein tumor necrosis factor–related apoptosis inducing ligand (TRAIL) and are currently preparing this genetically modified cell therapy for a phase 1/2a clinical trial in patients with metastatic lung cancer. To do this, we need to prepare a cryopreserved allogeneic MSCTRAIL cell bank for further expansion before patient delivery. The effects of cryopreservation on a genetically modified cell therapy product have not been clearly determined.

Methods

We tested different concentrations of dimethyl sulfoxide (DMSO) added to the human serum albumin ZENALB 4.5 and measured post-thaw cell viability, proliferation ability and differentiation characteristics. In addition, we examined the homing ability, TRAIL expression and cancer cell–killing capacities of cryopreserved genetically modified MSCs compared with fresh, continually cultured cells.

Results

We demonstrated that the post-thaw viability of MSCs in 5% DMSO (v/v) with 95% ZENALB 4.5 (v/v) is 85.7 ± 0.4%, which is comparable to that in conventional freezing media. We show that cryopreservation does not affect the long-term expression of TRAIL and that cryopreserved TRAIL-expressing MSCs exhibit similar levels of homing and, importantly, retain their potency in triggering cancer cell death.

Conclusions

This study shows that cryopreservation is unlikely to affect the therapeutic properties of MSCTRAIL and supports the generation of a cryopreserved master cell bank.