Novel methodology boosts MSC efficacy for articular cartilage restore

Researchers from the Essential Analytics for Manufacturing Customized-Medication (CAMP) Interdisciplinary Analysis Group (IRG) at 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 (NUSTEP), have developed a novel methodology to boost the flexibility of mesenchymal stromal cells (MSCs) to generate cartilage tissue by including ascorbic acid (AA) throughout MSC enlargement. The analysis additionally found that micro magnetic resonance relaxometry (µMRR), a novel course of analytical instrument (PAT) developed by SMART CAMP, can be utilized as a speedy, label-free process-monitoring instrument for the standard enlargement of MSCs.

Articular cartilage, a connective tissue that protects the bone ends in joints, can degenerate because of damage, age, or arthritis, resulting in vital joint ache and incapacity. Particularly in international locations – equivalent to Singapore – which have an lively, ageing inhabitants, articular cartilage degeneration is a rising ailment that impacts an rising variety of individuals. Autologous chondrocyte implantation is at present the one FDA-approved cell-based remedy for articular cartilage accidents, however it’s expensive, time-intensive, and requires a number of therapies. MSCs are a horny and promising various as they’ve proven good security profiles for transplantation. Nonetheless, scientific use of MSCs is proscribed because of inconsistent remedy outcomes arising from components equivalent to donor-to-donor variability, variation amongst cells throughout cell enlargement and unstandardized MSC manufacturing protocols.

The heterogeneity of MSCs can result in variations of their organic conduct and remedy 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. Due to this fact, improved processes are important to scale back cell heterogeneity whereas rising donor cell numbers with improved chondrogenic potential — the flexibility of MSCs to distinguish into cartilage cells to restore cartilage tissue — to pave the way in which for more practical and constant MSC-based therapies.

In a paper titled “Metabolic modulation to enhance MSC enlargement and therapeutic potential for articular cartilage restore”, printed within the scientific journal Stem Cell Analysis & Remedy, CAMP researchers detailed their growth of a priming technique to boost the enlargement of high quality MSCs by modifying the way in which cells make the most of vitality. 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 vitality that drives and helps many processes in dwelling cells. This means 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 reinforce their chondrogenic potential, they diverse 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 by which immature cells develop into mature cells with particular features, the researchers additional investigated its results throughout MSC enlargement.

The addition of AA to cell cultures for one passage throughout MSC enlargement and previous to initiation of differentiation was discovered to enhance chondrogenic differentiation, which is a vital high quality attribute (CQA) for higher articular cartilage restore. Longer-term AA remedy led to a greater than 300-fold enhance within the yield of MSCs with enhanced chondrogenic potential, diminished 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 sturdy shift in metabolic profile to OXPHOS. This metabolic change correlated with μMRR measurements, which helps determine novel CQAs that may very well be applied in MSC manufacturing for articular cartilage restore.

The analysis additionally demonstrates the potential of the method analytical instrument (PAT) 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 instrument for the enlargement of MSCs with AA supplementation. Initially used as a label-free malaria prognosis methodology because of the presence of paramagnetic hemozoin particles, μMRR was used within the analysis to detect senescence in MSCs. This speedy, 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 atmosphere — whereas enabling intermittent monitoring of a restricted lot measurement per manufacturing.

“Donor-to-donor variation, intrapopulation heterogeneity and mobile senescence have impeded the success of MSCs as a regular of care remedy for articular cartilage restore. Our analysis confirmed that AA supplementation throughout MSC enlargement can overcome these bottlenecks and improve MSC chondrogenic potential,” mentioned Ching Ann Tee, Senior Postdoctoral Affiliate at SMART CAMP, and first creator of the paper. 

By controlling metabolic circumstances equivalent to AA supplementation, coupled with CAMP’s course of analytical instruments equivalent to µMRR, the yield and high quality of cell remedy merchandise may very well be considerably elevated. This breakthrough may assist make MSC remedy a more practical and viable remedy possibility and supply requirements for bettering the manufacturing pipeline.” 

Ching Ann Tee, Senior Postdoctoral Affiliate, SMART CAMP

“This method of using metabolic modulation to enhance MSC chondrogenic potential may very well be tailored into related ideas for different therapeutic indications equivalent to osteogenic potential for bone restore or different varieties of stem cells. Implementing our findings in MSC manufacturing settings may very well be a big step ahead for sufferers with osteoarthritis and different joint illnesses, as we will effectively produce massive portions of high-quality MSCs with constant performance and allow the remedy of extra sufferers,” added Prof Laurie A. Boyer, Principal Investigator at SMART CAMP, Professor of Biology and Organic Engineering at MIT and corresponding creator of the paper.

The analysis is performed by SMART and supported by the Nationwide Analysis Basis (NRF) Singapore below its Campus for Analysis Excellence and Technological Enterprise (CREATE) programme.