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GMP-manufactured de...
GMP-manufactured density gradient media for optimized mesenchymal stromal/stem cell isolation and expansion
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Grisendi, Giulia (författare)
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- Annerén, Cecilia, 1972- (författare)
- Uppsala universitet,Institutionen för medicinsk biokemi och mikrobiologi
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Cafarelli, Luigi (författare)
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Sternieri, Rita (författare)
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Veronesi, Elena (författare)
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Cervo, Gian Luca (författare)
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Luminari, Stefano (författare)
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Maur, Michela (författare)
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Frassoldati, Antonio (författare)
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Palazzi, Giovanni (författare)
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Otsuro, satoru (författare)
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Franco, Bambi (författare)
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Paolucci, Paolo (författare)
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Pierfranco, Conte (författare)
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Horwitz, Edwin (författare)
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Dominici, Massimo (författare)
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(creator_code:org_t)
- Elsevier BV, 2010
- 2010
- Engelska.
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Ingår i: Cytotherapy. - : Elsevier BV. - 1465-3249 .- 1477-2566. ; 12:4, s. 466-477
- Relaterad länk:
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https://urn.kb.se/re...
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https://doi.org/10.3...
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Abstract
Ämnesord
Stäng
- BACKGROUND AIMS: Bone marrow (BM) mesenchymal stromal/stem cells (MSC) are therapeutic tools in regenerative medicine and oncology. MSC isolation is often performed starting from a separation step based on research-grade 1.077 g/mL density gradient media (DGM). However, MSC clinical application should require the introduction of good manufacturing practice (GMP) reagents. We took advantage of two novel GMP DGM with densities of 1.077 and 1.073 g/mL (Ficoll-Paque PREMIUM and Ficoll-Paque PREMIUM 1.073, respectively) to test whether these reagents could isolate MSC efficiently while simultaneously comparing their performance. METHODS: BM samples were processed using either 1.077 or 1.073 g/mL GMP DGM. BM mononucleated cell (MNC) fractions were analyzed for viability, immunophenotype, clonogenic potential, ex vivo expansion and differentiation potential. RESULTS: No differences were noticed in cell recovery and viability between the groups. Fluorescence-activated cell-sorting (FACS) analyzes on freshly isolated cells indicated that the 1.073 g/mL GMP DGM more efficiently depleted the CD45(+) fraction in comparison with 1.077 GMP DGM. Moreover, in the 1.073 group, fibroblastic colony-forming units (CFU-F) were 1.5 times higher and the final MSC yield 1.8 times increased after four passages. Both reagents isolated MSC with the expected phenotype; however, 1.073-isolated MSC showed a higher expression of CD90, CD146 and GD2. Additionally, MSC from both groups were capable of fully differentiating into bone, adipose cells and cartilage. CONCLUSIONS: Both GMP DGM enriched MSC from BM samples, suggesting that these reagents would be suitable for clinical-grade expansions. In addition, the density of 1.073 g/mL provides a significant advantage over 1.077 g/mL GMP DGM, impacting the quantity of MSC obtained and reducing the ex vivo expansion time for optimized cell-based clinical applications.
Ämnesord
- MEDICIN OCH HÄLSOVETENSKAP -- Medicinska och farmaceutiska grundvetenskaper -- Cell- och molekylärbiologi (hsv//swe)
- MEDICAL AND HEALTH SCIENCES -- Basic Medicine -- Cell and Molecular Biology (hsv//eng)
Nyckelord
- cell expansion
- density gradient media
- good manufacturing practice
- 1.073 g/mL
- Ficoll-Paque PREMIUM
- mesenchymal stromal/stem cells
- Morphology, cell biology, pathology
- Morfologi, cellbiologi, patologi
Publikations- och innehållstyp
- ref (ämneskategori)
- art (ämneskategori)
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Till lärosätets databas
- Av författaren/redakt...
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Grisendi, Giulia
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Annerén, Cecilia ...
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Cafarelli, Luigi
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Sternieri, Rita
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Veronesi, Elena
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Cervo, Gian Luca
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visa fler...
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Luminari, Stefan ...
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Maur, Michela
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Frassoldati, Ant ...
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Palazzi, Giovann ...
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Otsuro, satoru
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Franco, Bambi
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Paolucci, Paolo
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Pierfranco, Cont ...
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Horwitz, Edwin
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Dominici, Massim ...
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visa färre...
- Om ämnet
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- MEDICIN OCH HÄLSOVETENSKAP
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MEDICIN OCH HÄLS ...
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och Medicinska och f ...
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och Cell och molekyl ...
- Artiklar i publikationen
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Cytotherapy
- Av lärosätet
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Uppsala universitet