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



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.



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.



This study shows that cryopreservation is unlikely to affect the therapeutic properties of MSCTRAIL and supports the generation of a cryopreserved master cell bank.
Lungs for Living Research Centre, UCL Respiratory, Division of Medicine, University College London, London, United Kingdom.
Zhengqiang
Z
Yuan
Lungs for Living Research Centre, UCL Respiratory, Division of Medicine, University College London, London, United Kingdom.
Sofia Da Silva
Sda S
Lourenco
Lungs for Living Research Centre, UCL Respiratory, Division of Medicine, University College London, London, United Kingdom.
Elizabeth K
EK
Sage
Lungs for Living Research Centre, UCL Respiratory, Division of Medicine, University College London, London, United Kingdom.
Krishna K
KK
Kolluri
Centre for Cell, Gene & Tissue Therapy, Royal Free London National Health Services Foundation Trust & University College London, London, United Kingdom.
Mark W
MW
Lowdell
Lungs for Living Research Centre, UCL Respiratory, Division of Medicine, University College London, London, United Kingdom. Electronic address: s.janes@ucl.ac.uk.
Sam M
SM
Janes
D002454:Cell Differentiation; D045744:Cell Line, Tumor; D002465:Cell Movement; D049109:Cell Proliferation; D048430:Cell Shape; D002470:Cell Survival; D002478:Cells, Cultured; D015925:Cryopreservation / Q000379:methods*; D005615:Freezing; D006801:Humans; D045164:Mesenchymal Stem Cell Transplantation; D059630:Mesenchymal Stem Cells / Q000166:cytology* / Q000378:metabolism*; D009369:Neoplasms / Q000378:metabolism / Q000473:pathology / Q000628:therapy*; D019718:Receptors, CXCR4 / Q000378:metabolism; D053221:TNF-Related Apoptosis-Inducing Ligand / Q000378:metabolism*
D019718:Receptors, CXCR4; D053221:TNF-Related Apoptosis-Inducing Ligand
DMSO; MSC; ZENALB 4.5; apoptosis; chemokine; cryopreservation