Quality Control of Umbilical Cord Mesenchymal Stem Cells for the Treatment of Systemic Lupus Erythematosus

摘要

Objective: To isolate and cultivate umbilical cord mesenchymal stem cells (UC-MSC) suitable for the treatment of systemic lupus erythematosus (SLE). Methods: MSCs were obtained from umbilical cord by adherent method. After subculture, UC-MSC was amplified by serum-free culture system. Through flow cytometry and induced differentiation technology, stem cell content, total number of cells and cell viability were detected, and samples were taken for bacterial detection, endotoxin detection and mycoplasma detection to meet the quality standards before application. To establish sound technical procedures and quality standards for the production, packaging, storage and transportation of umbilical cord mesenchymal stem cells; complete the quality control system plan, BALB/C mice were selected and pristane was used for modeling. BALB/C mice were randomly divided into control group, model group and treatment group. The treatment group was given UC-MSC (4 × 105 cells/ml), 0.2 ml. Antidouble-stranded DNA (ds-DNA) antibodies, systemic lupus erythematous Activity score (SLEDAI), and routine blood tests were performed. The safety of umbilical cord mesenchymal stem cells was evaluated by tumorigenicity test and toxicity test in nude mice. Results and Conclusion: After technical modification and upgrading, the serum-free culture of UC-MSCs grew adherently, mainly in the form of typical fibroblasts, which could be transmitted over 15 generations. The fifth generation MSCs showed high expression of CD73, CD105, CD90, CD44 and low expression of CD34, CD45 and HLA-DR. Under different induction conditions, umbilical cord MSCs could be induced to differentiate into osteoblasts and chondrocytes. After 30 days of UC-MSC administration, the level of anti-DS -DNA antibody in the treatment group was significantly reduced, with significant differences compared with the control group. The disease activity score was significantly reduced, and indicators such as proteinuria, serum creatinine and urea nitrogen were improved, with statistically significant differences compared with the model group (P 0.05). This study established a quality management system for the industrial production of UC-MSC, verified the safety and effectiveness of UC-MSC in the treatment of systemic lupus erythematosus, and provided theoretical basis and technical support for subsequent clinical trials of UC-MSC in the treatment of systemic lupus erythematosus.