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

• Published in: Journal of translational medicine Vol. 14; no. 1; p. 319
• Main Authors: Haack-Sørensen, Mandana, Follin, Bjarke, Juhl, Morten, Brorsen, Sonja K, Søndergaard, Rebekka H, Kastrup, Jens, Ekblond, Annette
• Format: Journal Article
• Language: English
• Published: England BioMed Central Ltd 16-11-2016; BioMed Central; BMC
• Subjects: Adipose derived stromal cells; Adipose Tissue - cytology; Adipose tissues; Adult; Aged; Analysis; Automation; Bioreactors; Cell Count; Cell culture; Cell Culture Techniques - methods; Cell Differentiation; Cell Lineage; Cell Proliferation; Cell Survival; Cells, Cultured; Coating; Comparative analysis; Cryoprecipitate; Female; Flow Cytometry; Gene expression; Health aspects; Humans; Immunophenotyping; Medical care, Cost of; Mesenchymal stem cell; Middle Aged; Phenotype; Stem cells; Storage; Stromal Cells - cytology; Denmark; Mesenchymal stem cell; Cell culture; Bioreactor; Storage; Clinical application; Cell expansion; Adipose derived stromal cells; Coating; Cryoprecipitate
• ISSN: 1479-5876; 1479-5876
• DOI: 10.1186/s12967-016-1080-9

Adipose 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.
Stromal 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.
The 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 × 10
SVF cells loaded into a Quantum system yielded 8.96 × 10
ASCs P0, while 4.5 × 10
SVF cells seeded per T75 flask yielded an average of 2.37 × 10
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.
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.