Towards an advanced therapy medicinal product based on mesenchymal stromal cells isolated from the umbilical cord tissue: quality and safety data

2.50
Hdl Handle:
http://hdl.handle.net/10033/621011
Title:
Towards an advanced therapy medicinal product based on mesenchymal stromal cells isolated from the umbilical cord tissue: quality and safety data
Authors:
Martins, José P; Santos, Jorge M; Almeida, Joana M d; Filipe, Mariana A; de Almeida, Mariana V T; Almeida, Sílvia C C; Água-Doce, Ana; Varela, Alexandre; Gilljam, Mari; Stellan, Birgitta; Pohl, Susanne; Dittmar, Kurt; Lindenmaier, Werner; Alici, Evren; Graça, Luís; Cruz, Pedro E; Cruz, Helder J; Bárcia, Rita N
Abstract:
Abstract Introduction Standardization of mesenchymal stromal cells (MSCs) manufacturing is urgently needed to enable translational activities and ultimately facilitate comparison of clinical trial results. In this work we describe the adaptation of a proprietary method for isolation of a specific umbilical cord tissue-derived population of MSCs, herein designated by its registered trademark as UCX®, towards the production of an advanced therapy medicinal product (ATMP). Methods The adaptation focused on different stages of production, from cell isolation steps to cell culturing and cryopreservation. The origin and quality of materials and reagents were considered and steps for avoiding microbiological and endotoxin contamination of the final cell product were implemented. Cell isolation efficiency, MSCs surface markers and genetic profiles, originating from the use of different medium supplements, were compared. The ATMP-compliant UCX® product was also cryopreserved avoiding the use of dimethyl sulfoxide, an added benefit for the use of these cells as an ATMP. Cells were analyzed for expansion capacity and longevity. The final cell product was further characterized by flow cytometry, differentiation potential, and tested for contaminants at various passages. Finally, genetic stability and immune properties were also analyzed. Results The isolation efficiency of UCX® was not affected by the introduction of clinical grade enzymes. Furthermore, isolation efficiencies and phenotype analyses revealed advantages in the use of human serum in cell culture as opposed to human platelet lysate. Initial decontamination of the tissue followed by the use of mycoplasma- and endotoxin-free materials and reagents in cell isolation and subsequent culture, enabled the removal of antibiotics during cell expansion. UCX®-ATMP maintained a significant expansion potential of 2.5 population doublings per week up to passage 15 (P15). They were also efficiently cryopreserved in a DMSO-free cryoprotectant medium with approximately 100% recovery and 98% viability post-thaw. Additionally, UCX®-ATMP were genetically stable upon expansion (up to P15) and maintained their immunomodulatory properties. Conclusions We have successfully adapted a method to consistently isolate, expand and cryopreserve a well-characterized population of human umbilical cord tissue-derived MSCs (UCX®), in order to obtain a cell product that is compliant with cell therapy. Here, we present quality and safety data that support the use of the UCX® as an ATMP, according to existing international guidelines.
Citation:
Stem Cell Research & Therapy. 2014 Jan 17;5(1):9
Issue Date:
17-Jan-2014
URI:
http://dx.doi.org/10.1186/scrt398; http://hdl.handle.net/10033/621011
Type:
Journal Article
Appears in Collections:
Publications of Dept. Gene Regulation and Differentiation (RDIF)

Full metadata record

DC FieldValue Language
dc.contributor.authorMartins, José Pen
dc.contributor.authorSantos, Jorge Men
dc.contributor.authorAlmeida, Joana M den
dc.contributor.authorFilipe, Mariana Aen
dc.contributor.authorde Almeida, Mariana V Ten
dc.contributor.authorAlmeida, Sílvia C Cen
dc.contributor.authorÁgua-Doce, Anaen
dc.contributor.authorVarela, Alexandreen
dc.contributor.authorGilljam, Marien
dc.contributor.authorStellan, Birgittaen
dc.contributor.authorPohl, Susanneen
dc.contributor.authorDittmar, Kurten
dc.contributor.authorLindenmaier, Werneren
dc.contributor.authorAlici, Evrenen
dc.contributor.authorGraça, Luísen
dc.contributor.authorCruz, Pedro Een
dc.contributor.authorCruz, Helder Jen
dc.contributor.authorBárcia, Rita Nen
dc.date.accessioned2017-07-13T11:59:23Z-
dc.date.available2017-07-13T11:59:23Z-
dc.date.issued2014-01-17en
dc.identifier.citationStem Cell Research & Therapy. 2014 Jan 17;5(1):9en
dc.identifier.urihttp://dx.doi.org/10.1186/scrt398en
dc.identifier.urihttp://hdl.handle.net/10033/621011-
dc.description.abstractAbstract Introduction Standardization of mesenchymal stromal cells (MSCs) manufacturing is urgently needed to enable translational activities and ultimately facilitate comparison of clinical trial results. In this work we describe the adaptation of a proprietary method for isolation of a specific umbilical cord tissue-derived population of MSCs, herein designated by its registered trademark as UCX®, towards the production of an advanced therapy medicinal product (ATMP). Methods The adaptation focused on different stages of production, from cell isolation steps to cell culturing and cryopreservation. The origin and quality of materials and reagents were considered and steps for avoiding microbiological and endotoxin contamination of the final cell product were implemented. Cell isolation efficiency, MSCs surface markers and genetic profiles, originating from the use of different medium supplements, were compared. The ATMP-compliant UCX® product was also cryopreserved avoiding the use of dimethyl sulfoxide, an added benefit for the use of these cells as an ATMP. Cells were analyzed for expansion capacity and longevity. The final cell product was further characterized by flow cytometry, differentiation potential, and tested for contaminants at various passages. Finally, genetic stability and immune properties were also analyzed. Results The isolation efficiency of UCX® was not affected by the introduction of clinical grade enzymes. Furthermore, isolation efficiencies and phenotype analyses revealed advantages in the use of human serum in cell culture as opposed to human platelet lysate. Initial decontamination of the tissue followed by the use of mycoplasma- and endotoxin-free materials and reagents in cell isolation and subsequent culture, enabled the removal of antibiotics during cell expansion. UCX®-ATMP maintained a significant expansion potential of 2.5 population doublings per week up to passage 15 (P15). They were also efficiently cryopreserved in a DMSO-free cryoprotectant medium with approximately 100% recovery and 98% viability post-thaw. Additionally, UCX®-ATMP were genetically stable upon expansion (up to P15) and maintained their immunomodulatory properties. Conclusions We have successfully adapted a method to consistently isolate, expand and cryopreserve a well-characterized population of human umbilical cord tissue-derived MSCs (UCX®), in order to obtain a cell product that is compliant with cell therapy. Here, we present quality and safety data that support the use of the UCX® as an ATMP, according to existing international guidelines.en
dc.titleTowards an advanced therapy medicinal product based on mesenchymal stromal cells isolated from the umbilical cord tissue: quality and safety dataen
dc.typeJournal Articleen
dc.language.rfc3066enen
dc.rights.holderMartins et al.; licensee BioMed Central Ltd.en
dc.date.updated2015-09-04T08:21:57Zen
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