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Culture expansion of adipose derived stromal cells. A closed automated Quantum Cell Expansion System compared with manual flask-based culture
- Source :
- Haack-Sørensen, M, Follin, B, Juhl, M, Brorsen, S K, Søndergaard, R, Kastrup, J & Ekblond, A 2016, ' Culture expansion of adipose derived stromal cells. A closed automated Quantum Cell Expansion System compared with manual flask-based culture ', Journal of Translational Medicine, vol. 14, no. 1, 319 . https://doi.org/10.1186/s12967-016-1080-9, Journal of Translational Medicine, Vol 14, Iss 1, Pp 1-11 (2016), Journal of Translational Medicine
- Publication Year :
- 2016
-
Abstract
- Background 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. Methods 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. Results 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 × 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 material The online version of this article (doi:10.1186/s12967-016-1080-9) contains supplementary material, which is available to authorized users.
- Subjects :
- 0301 basic medicine
Pathology
Cell
Cell Culture Techniques
Adipose tissue
lcsh:Medicine
Storage
Cell Count
Coating
Automation
Cell expansion
Bioreactors
Cells, Cultured
Mesenchymal stem cell
Medicine(all)
Cell Differentiation
General Medicine
Middle Aged
Stromal vascular fraction
Flow Cytometry
Clinical application
Cell biology
Phenotype
medicine.anatomical_structure
Adipose Tissue
Female
Adult
medicine.medical_specialty
Stromal cell
Cell Survival
Cryoprecipitate
Bioreactor
Biology
General Biochemistry, Genetics and Molecular Biology
Immunophenotyping
03 medical and health sciences
Laboratory flask
medicine
Humans
Cell Lineage
Aged
Cell Proliferation
Biochemistry, Genetics and Molecular Biology(all)
Research
lcsh:R
equipment and supplies
030104 developmental biology
Cell culture
Stromal Cells
Adipose derived stromal cells
Fetal bovine serum
Subjects
Details
- Language :
- English
- Database :
- OpenAIRE
- Journal :
- Haack-Sørensen, M, Follin, B, Juhl, M, Brorsen, S K, Søndergaard, R, Kastrup, J & Ekblond, A 2016, ' Culture expansion of adipose derived stromal cells. A closed automated Quantum Cell Expansion System compared with manual flask-based culture ', Journal of Translational Medicine, vol. 14, no. 1, 319 . https://doi.org/10.1186/s12967-016-1080-9, Journal of Translational Medicine, Vol 14, Iss 1, Pp 1-11 (2016), Journal of Translational Medicine
- Accession number :
- edsair.doi.dedup.....30ca5f34aab82642e80c2d7e9ed8cd05
- Full Text :
- https://doi.org/10.1186/s12967-016-1080-9