Culture expansion of adipose derived stromal cells. A closed automated Quantum Cell Expansion System compared with manual flask-based culture

Abstract

BackgroundAdipose derived stromal cells (ASCs) are a rich and convenient source of cells for clinical regenerative therapeutic approaches. However, applications of ASCs often require cell expansion to reach the needed dose. In this study, cultivation of ASCs from stromal vascular fraction (SVF) over two passages in the automated and functionally closed Quantum Cell Expansion System (Quantum system) is compared with traditional manual cultivation.MethodsStromal vascular fraction was isolated from abdominal fat, suspended in α-MEM supplemented with 10% Fetal Bovine Serum and seeded into either T75 flasks or a Quantum system that had been coated with cryoprecipitate. The cultivation of ASCs from SVF was performed in 3 ways: flask to flask; flask to Quantum system; and Quantum system to Quantum system. In all cases, quality controls were conducted for sterility, mycoplasmas, and endotoxins, in addition to the assessment of cell counts, viability, immunophenotype, and differentiation potential.ResultsThe viability of ASCs passage 0 (P0) and P1 was above 96%, regardless of cultivation in flasks or Quantum system. Expression of surface markers and differentiation potential was consistent with ISCT/IFATS standards for the ASC phenotype. Sterility, mycoplasma, and endotoxin tests were consistently negative. An average of 8.0 × 107 SVF cells loaded into a Quantum system yielded 8.96 × 107 ASCs P0, while 4.5 × 106 SVF cells seeded per T75 flask yielded an average of 2.37 × 106 ASCs—less than the number of SVF cells seeded. ASCs P1 expanded in the Quantum system demonstrated a population doubling (PD) around 2.2 regardless of whether P0 was previously cultured in flasks or Quantum, while ASCs P1 in flasks only reached a PD of 1.0. Conclusion: Manufacturing of ASCs in a Quantum system enhances ASC expansion rate and yield significantly relative to manual processing in T-flasks, while maintaining the purity and quality essential to safe and robust cell production. Notably, the use of the Quantum system entails significantly reduced working hours and thereby costs.Electronic supplementary materialThe online version of this article (doi:10.1186/s12967-016-1080-9) contains supplementary material, which is available to authorized users.

Cryopreserved Off-the-Shelf Allogeneic Adipose-Derived Stromal Cells for Therapy in Patients with Ischemic Heart Disease and Heart Failure—A Safety Study

“…The overall aim was to establish a logistically and clinically applicable off‐the shelf stem cell treatment strategy for patients. We changed from autologous to allogeneic cells, and we implemented a more standardized and reproducible stem cell production platform using semi‐automated closed bioreactor systems instead of flasks and human platelet lysate instead of fetal bovine serum, as well as cryo storage of a ready‐to‐use product .…”

“…The overall aim was to establish a logistically and clinically applicable off-the shelf stem cell treatment strategy for patients. We changed from autologous to allogeneic cells, and we implemented a more standardized and reproducible stem cell production platform using semi-automated closed bioreactor systems instead of flasks and human platelet lysate instead of fetal bovine serum, as well as cryo storage of a ready-to-use product [7][8][9][10][11][12][13][14][15][16][17][18][19]. We have in the present study demonstrated that it is feasible to generate a reproducible, standardized, and clinically applicable cell product from healthy donors that is safe and has a very promising efficacy profile.…”

Rationale and design of the European multicentre study on Stem Cell therapy in IschEmic Non‐treatable Cardiac diseasE (SCIENCE)

“…Purity, differentiation capacity (adipose, bone, cartilage), and full phenotyping consistent with ISCT/IFATS standards for the ASC phenotype as well as genomic stability has been determined during manufacturing process development . Stability studies have been established, and currently 24‐month stability demonstrating sterility, viability, recovery, ASC immunophenotype, and cell potency as defined by in vitro cell adherence and proliferation, after thawing of the final product, has been documented and approved by competent authorities.…”

Bioprocess Development for Human Mesenchymal Stem Cell Therapy Products

“…The shear forces in fixed bed reactors are low (~0.5 × 10 −5 N/cm 2 ) and consistent throughout the reactor with no peaks near the impeller; the shear forces also remain constant at all scales [81]. The in vitro expansion of MSCs has been reported in several types of fixed bed reactors [82][83][84][85]. One of the major drawbacks of fixed bed and other reactor types compared to stirred tank reactors is the challenge of efficient harvesting.…”