Study of the stability of packaging and storage conditions of human mesenchymal stem cell for intra-arterial clinical application in patient with critical limb ischemia.
Critical limb ischemia (CLI) is associated with significant morbidity and mortality. In this study, we developed and characterized an intra-arterial cell suspension containing human mesenchymal stem cells (hMSCs) for the treatment of CLI. Equally, the stability of cells was studied in order to evaluate the optimal conditions of storage that guarantee the viability from cell processing to the administration phase. Effects of various factors, including excipients, storage temperature and time were evaluated to analyze the survival of hMSCs in the finished medicinal product. The viability of hMSCs in different packaging media was studied for 60 h at 4 °C. The best medium to maintain hMSCs viability was then selected to test storage conditions (4, 8, 25 and 37 °C; 60 h). The results showed that at 4 °C the viability was maintained above 80% for 48 h, at 8 °C decreased slightly, whereas at room temperature and 37 °C decreased drastically. Its biocompatibility was assessed by cell morphology and cell viability assays. During stability study, the stored cells did not show any change in their phenotypic or genotypic characteristics and physicochemical properties remained constant, the ability to differentiate into adipocytes and osteocytes and sterility requirements were also unaltered. Finally, our paper proposes a packing media composed of albumin 20%, glucose 5% and Ringer's lactate at a concentration of 1×10(6) cells/mL, which must be stored at 4 °C as the most suitable to maintain cell viability (>80%) and without altering their characteristics for more than 48 h.
Andalusian Centre for Molecular Biology and Regenerative Medicine (CABIMER), Seville, Spain; Department of Pharmacy and Pharmaceutical Technology, Faculty of Pharmacy, University of Granada, Spain.
Patricia
P
Gálvez-Martín
Andalusian Centre for Molecular Biology and Regenerative Medicine (CABIMER), Seville, Spain; Centre for Network Biomedical Research on Diabetes and Associated Metabolic Diseases (CIBERDEM), Barcelona, Spain.
Abdelkrim
A
Hmadcha
Andalusian Centre for Molecular Biology and Regenerative Medicine (CABIMER), Seville, Spain; Centre for Network Biomedical Research on Diabetes and Associated Metabolic Diseases (CIBERDEM), Barcelona, Spain.
Bernat
B
Soria
Department of Pharmacy and Pharmaceutical Technology, Faculty of Pharmacy, University of Barcelona, Spain.
Ana C
AC
Calpena-Campmany
Department of Pharmacy and Pharmaceutical Technology, Faculty of Pharmacy, University of Granada, Spain. Electronic address: beatrizclares@ugr.es.
Beatriz
B
Clares-Naveros
D002478:Cells, Cultured; D003925:Diabetic Angiopathies / Q000097:blood / Q000628:therapy*; D004349:Drug Packaging / Q000592:standards; D004355:Drug Stability; D004356:Drug Storage / Q000592:standards; D006801:Humans; D007269:Injections, Intra-Arterial; D045164:Mesenchymal Stem Cell Transplantation / Q000379:methods / Q000592:standards*; D059630:Mesenchymal Stem Cells / Q000737:chemistry* / Q000502:physiology; D014015:Tissue Banks / Q000592:standards
Adipose mesenchymal stem cell; Critical limb ischemia; Injectable; Intra-arterial; Packaging; Storage