Researchers from the Essential Analytics for Manufacturing Personalised-Drugs (CAMP) interdisciplinary analysis group on the Singapore-MIT Alliance for Analysis and Expertise (SMART), MIT’s analysis enterprise in Singapore, alongside collaborators from the Nationwide College of Singapore Tissue Engineering Programme, have developed a novel methodology to boost the power of mesenchymal stromal cells (MSCs) to generate cartilage tissue by including ascorbic acid throughout MSC growth. The analysis additionally found that micro-magnetic resonance relaxometry (µMRR), a novel course of analytical device developed by SMART CAMP, can be utilized as a fast, label-free process-monitoring device for the standard growth of MSCs.
Articular cartilage, a connective tissue that protects the bone ends in joints, can degenerate because of harm, age, or arthritis, resulting in important joint ache and incapacity. Particularly in nations — reminiscent of Singapore — which have an energetic, getting older inhabitants, articular cartilage degeneration is a rising ailment that impacts an growing variety of individuals. Autologous chondrocyte implantation is at the moment the one Meals and Drug Administration-approved cell-based remedy for articular cartilage accidents, however it’s pricey, time-intensive, and requires a number of remedies. MSCs are a beautiful and promising various as they’ve proven good security profiles for transplantation. Nonetheless, medical use of MSCs is proscribed because of inconsistent therapy outcomes arising from elements reminiscent of donor-to-donor variability, variation amongst cells throughout cell growth, and non-standardized MSC manufacturing protocols.
The heterogeneity of MSCs can result in variations of their organic conduct and therapy outcomes. Whereas large-scale MSC expansions are required to acquire a therapeutically related variety of cells for implantation, this course of can introduce cell heterogeneity. Subsequently, improved processes are important to cut back cell heterogeneity whereas growing donor cell numbers with improved chondrogenic potential — the power of MSCs to distinguish into cartilage cells to restore cartilage tissue — to pave the best way for simpler and constant MSC-based therapies.
In a paper titled “Metabolic modulation to enhance MSC growth and therapeutic potential for articular cartilage restore,” printed within the scientific journal Stem Cell Analysis and Remedy, CAMP researchers detailed their growth of a priming technique to boost the growth of high quality MSCs by modifying the best way cells make the most of power. The analysis findings have proven a optimistic correlation between chondrogenic potential and oxidative phosphorylation (OXPHOS), a course of that harnesses the discount of oxygen to create adenosine triphosphate — a supply of power that drives and helps many processes in dwelling cells. This implies that manipulating MSC metabolism is a promising technique for enhancing chondrogenic potential.
Utilizing novel PATs developed by CAMP, the researchers explored the potential of metabolic modulation in each short- and long-term harvesting and reseeding of cells. To boost their chondrogenic potential, they assorted the nutrient composition, together with glucose, pyruvate, glutamine, and ascorbic acid (AA). As AA is reported to assist OXPHOS and its optimistic affect on chondrogenic potential throughout differentiation — a course of wherein immature cells turn into mature cells with particular capabilities — the researchers additional investigated its results throughout MSC growth.
The addition of AA to cell cultures for one passage throughout MSC growth and previous to initiation of differentiation was discovered to enhance chondrogenic differentiation, which is a crucial high quality attribute (CQA) for higher articular cartilage restore. Longer-term AA therapy led to a greater than 300-fold enhance within the yield of MSCs with enhanced chondrogenic potential, and lowered cell heterogeneity and cell senescence — a course of by which a cell ages and completely stops dividing however doesn’t die — when in comparison with untreated cells. AA-treated MSCs with improved chondrogenic potential confirmed a strong shift in metabolic profile to OXPHOS. This metabolic change correlated with μMRR measurements, which helps establish novel CQAs that might be carried out in MSC manufacturing for articular cartilage restore.
The analysis additionally demonstrates the potential of the method analytical device developed by CAMP, micromagnetic resonance relaxometry (μMRR) — a miniature benchtop machine that employs magnetic resonance imaging (MRI) imaging on a microscopic scale — as a process-monitoring device for the growth of MSCs with AA supplementation. Initially used as a label-free malaria analysis methodology because of the presence of paramagnetic hemozoin particles, μMRR was used within the analysis to detect senescence in MSCs. This fast, label-free methodology requires solely a small variety of cells for analysis, which permits for MSC remedy manufacturing in closed methods — a system for safeguarding pharmaceutical merchandise by decreasing contamination dangers from the exterior surroundings — whereas enabling intermittent monitoring of a restricted lot dimension per manufacturing.
“Donor-to-donor variation, intrapopulation heterogeneity, and mobile senescence have impeded the success of MSCs as an ordinary of care remedy for articular cartilage restore. Our analysis confirmed that AA supplementation throughout MSC growth can overcome these bottlenecks and improve MSC chondrogenic potential,” says Ching Ann Tee, senior postdoc at SMART CAMP and first writer of the paper. “By controlling metabolic situations reminiscent of AA supplementation, coupled with CAMP’s course of analytical instruments reminiscent of µMRR, the yield and high quality of cell remedy merchandise might be considerably elevated. This breakthrough might assist make MSC remedy a simpler and viable therapy choice and supply requirements for bettering the manufacturing pipeline.”
“This method of using metabolic modulation to enhance MSC chondrogenic potential might be tailored into related ideas for different therapeutic indications, reminiscent of osteogenic potential for bone restore or different sorts of stem cells. Implementing our findings in MSC manufacturing settings might be a big step ahead for sufferers with osteoarthritis and different joint ailments, as we will effectively produce massive portions of high-quality MSCs with constant performance and allow the therapy of extra sufferers,” provides Professor Laurie A. Boyer, principal investigator at SMART CAMP, professor of biology and organic engineering at MIT, and corresponding writer of the paper.
The analysis is performed by SMART and supported by the Nationwide Analysis Basis Singapore beneath its Campus for Analysis Excellence and Technological Enterprise program.
