Large-scale bioreactor expansion of tumor-infiltrating lymphocytes.
BACKGROUND
The aim of this study was to evaluate an improved technique for expansion of tumor-infiltrating lymphocytes (TILs) based on the WAVE Bioreactor system with perfusion and tube-welding techniques. Our hypothesis was that the bioreactor would allow for optimized provision of nutrients and removal of spent media while minimizing culture volumes. These refinements might lead to a better quality of expanded cells with lower amounts of exhausted cells compared to static expansions in culture bags.


METHODS
Tumor-infiltrating lymphocytes from 4 melanoma patients were expanded and compared in parallel using either the WAVE Bioreactor 2/10 System or traditional static culture methods. The parameters viability, final cell number, phenotype and effector function were measured.


RESULTS
Our results show that the bioreactor system with perfusion is suitable for large-scale expansion of tumor-infiltrating lymphocytes and allows for higher cell densities and absolute cell numbers as compared to static culture conditions. Phenotypic characteristics of TILs were compared pre and post expansion and showed no consistent difference between the two expansion methods. TILs harvested had the phenotype and function corresponding to intermediate to late effector cells. The system allows one technician to operate several bioreactors simultaneously, thereby reducing the labor for one expansion to approximately 1/3 compared to static expansion.


CONCLUSIONS
The WAVE Bioreactor system is suitable for large-scale expansion of TILs. Due to constant perfusion of fresh media and removal of spent media much higher cell densities were achieved while the culture volume and the glucose and glutamine levels were kept constant. Expansion of TILs in the bioreactor system represents a labor- and cost-effective method to reach large numbers of T cells for adoptive cell transfer therapy in the clinic.


CONCLUSIONS
The system presented herein offers an effective alternative to large-scale production of cell products for clinical use while meeting requirements of therapeutic cell quantities and qualities of current protocols for treatment of malignant melanoma.
Department of Oncology, Radiology and Clinical Immunology, Division of Clinical Immunology, Rudbeck Laboratory, Uppsala University, Sweden. arian.sadeghi@klinimm.uu.se
Arian
A
Sadeghi
Linnea
L
Pauler
Cecilia
C
Annerén
Andrew
A
Friberg
Daniel
D
Brandhorst
Olle
O
Korsgren
Thomas H
TH
Tötterman
D024221:Antineoplastic Protocols; D019149:Bioreactors; D002452:Cell Count; D018929:Cell Culture Techniques; D045744:Cell Line, Tumor; D049109:Cell Proliferation; D002470:Cell Survival; D018920:Coculture Techniques; D003362:Cost-Benefit Analysis; D006801:Humans; D016130:Immunophenotyping; D016219:Immunotherapy, Adoptive / Q000191:economics / Q000295:instrumentation / Q000379:methods; D007371:Interferon-gamma / Q000378:metabolism; D008213:Lymphocyte Activation; D016246:Lymphocytes, Tumor-Infiltrating / Q000276:immunology / Q000378:metabolism* / Q000473:pathology; D008545:Melanoma / Q000276:immunology / Q000473:pathology / Q000628:therapy*; D012878:Skin Neoplasms / Q000276:immunology / Q000473:pathology / Q000628:therapy*
D007371:Interferon-gamma