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New advanced therapy medicinal product for cartilage defect treatment with extemporaneous association of a scaffold and Wharton's jelly mesenchymal stromal cells
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International audience. n the framework of an ANR project obtained in 2015, we propose to develop a new advancedtherapy medicinal product for cartilage defect treatment with extemporaneous association of ascaffold and Wharton’s jelly mesenchymal stromal cells (WJ-MSC).Due to their capacity of proliferation and differentiation, MSC appear to be currently one of themost promising ways of obtaining cartilage cells. The conjunctive tissue of the umbilical cordor Wharton’s jelly is an abundant and promising source of MSC for clinical applications. Theypresent immunomodulatory properties and have a higher proliferation potential than MSC fromadult tissues. All these advantages enable this source to represent a virtually inexhaustiblesource of stem cells especially for allogeneic tissue engineering therapies.The first part of this work made it possible to perform a technological transfer from researchtowards the clinic. The transfer consisted in adjusting MSC isolation, production andconservation methods developed by research lab in clinical grade conditions.Eight umbilical cords were obtained after the signing of an informed consent form by pregnantmothers. In parallel, for each umbilical cord collected, well identified obstetric factors wererecorded. We developed a fast, simple and efficient explant method for the isolation of MSCfrom human umbilical cords requiring minimal manipulation and thus reducing the risk ofcontamination. MSC isolation, production and conservation were performed in a controlledatmosphere zone. All reagents and materials were well-defined and controlled. Cell culture wasperformed in culture containers (CellSTACK®, Macopharma) in a closed system. MSC werecultivated in hypoxia until passage 2 and then frozen. Quality controls were carried out duringproduction: serological and microbiological controls, cell number, viability, clonogeniccapacity, immunophenotype, mesodermic differentiation potential and karyotype.This isolation method showed a 100% success rate. We were able to obtain a large number ofviable and secured cells with an average number of 50.106 cells per CellSTACK and a doublingtime of about 1.5 days. During the culture, WJ-MSC exhibited a sustained clonogenic capacityand a specific mesenchymal phenotype.These results made it possible to assess the feasibility and to validate the clinical gradeamplification technique as well as to obtain precisely characterized and secured WJ-MSC.
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