Manufacturing Of Cellular Products Research Articles

A risk assessment study of SARS-CoV-2 propagation in the manufacturing of cellular products.

The potential infection of cellular therapies by SARS-CoV-2 present high risks, as the target patients for these treatments are often immunocompromised or have chronic diseases associated with a higher risk of serious illness and death by COVID-19. The multicellular tropism of this virus presents challenges for the manufacturing of cell therapies, whereby the material could potentially become infected at the source or during cell processing. In this review we assess the risk of a SARS-CoV-2 propagation in cell types used to date in cellular therapies. Altogether, the risk of SARS-CoV-2 contamination of cellular products remains low. This risk should be evaluated on an individual basis, considering ACE2 and TMPRSS2 expression, existing literature regarding the susceptibility to infection, and single cell RNA sequencing data of COVID-19 patients. This analysis should ideally be performed for both the cells being manufactured and the cells used to produce the vector to ensure patient safety.

Membrane Applications in Autologous Cell Therapy.

Stem cell and cell therapies, particularly autologous cell therapies, are becoming a common practice. However, in order for these technologies to achieve wide-scale clinical application, the prohibitively high cost associated with these therapies must be addressed through creative engineering. Membranes can be a disruptive technology to reshape the bioprocessing and manufacture of cellular products and significantly reduce the cost of autologous cell therapies. Examples of successful membrane applications include expansions of CAR-T cells, various human stem cells, and production of extracellular vesicles (EVs) using hollow fibre membrane bioreactors. Novel membranes with tailored functions and surface properties and novel membrane modules that can accommodate the changing needs for surface area and transport properties are to be developed to fulfil this key role.

Cells to the Rescue: Emerging Cell-Based Treatment Approaches for NMOSD and MOGAD.

Cell-based therapies are gaining momentum as promising treatments for rare neurological autoimmune diseases, including neuromyelitis optica spectrum disorders and myelin oligodendrocyte glycoprotein antibody-associated disease. The development of targeted cell therapies is hampered by the lack of adequate animal models that mirror the human disease. Most cell-based treatments, including HSCT, CAR-T cell, tolerogenic dendritic cell and mesenchymal stem cell treatment have entered early stage clinical trials or have been used as rescue treatment in treatment-refractory cases. The development of antigen-specific cell-based immunotherapies for autoimmune diseases is slowed down by the rarity of the diseases, the lack of surrogate outcomes and biomarkers that are able to predict long-term outcomes and/or therapy effectiveness as well as challenges in the manufacturing of cellular products. These challenges are likely to be overcome by future research.

Evaluation of human platelet lysate (HPL) as a substitute for foetal bovine serum (FBS) and recombinant proteins for the growth and maintenance of bone marrow derived mesenchymal stromal cells (BM-MSC)

Aim To evaluate the potential of using HPL produced in Australia as a substitute for FBS during in vitro culture of BM-MSC. Background The replacement of FBS with an alternate serum source during the manufacture of cellular products for human therapeutics should minimise cost and exposure to the potential risk of animal products. Human AB sera serves as the current gold standard in clinical grade manufacture of cell therapies but suffers from high cost and limited supply. Here, we propose the use of HPL as an FBS replacement. Methods We compared BM-MSC cultured in FBS or HPL by assessing their growth, viability and phenotype by flow cytometry at P3 and P4. Three batches of HPL made from either apheresis or pooled platelet concentrates after expiry were tested. BM-MSC from a single donor were seeded at 1.8 × 103/cm2 in culture flasks pre-coated with 0.0001% human fibronectin, and passaged at 80% confluency. Cells were cultured in DMEM supplemented with 5% apheresis HPL, 10% pooled HPL or 10% Commercial HPL, compared to the Control media (10% FBS, 5 ng/mL PDGF, and 5 ng/mL EGF). As preliminary analysis revealed minimal variability between batches of HPL, we focused the remaining experiments on one batch of each type of HPL. Results The phenotype of all BM-MSC at P3 and P4 satisfied established MSC criteria, that is, >98% expression of CD73, CD90 and CD105, and lack of expression (i.e. 0.05 by one way ANOVA). In contrast, pooled HPL did not support BM-MSC growth beyond P3. This may be due to differences in the characteristics of apheresis and pooled HPL. Conclusion The growth and phenotype of BM-MSC cultured in either apheresis HPL or commercial HPL were comparable, suggesting apheresis HPL may prove a suitable alternative to FBS. A simpler preparation protocol of HPL for the end users will increase the likelihood of introducing HPL into current laboratory practice. The potential of pooled HPL warrants further investigation.

Manufacture of tumor- and virus-specific T lymphocytes for adoptive cell therapies.

Adoptive transfer of tumor-infiltrating lymphocytes (TILs) and genetically engineered T lymphocytes expressing chimeric antigen receptors (CARs) or conventional alpha/beta T-cell receptors (TCRs), collectively termed adoptive cell therapy (ACT), is an emerging novel strategy to treat cancer patients. Application of ACT has been constrained by the ability to isolate and expand functional tumor-reactive T cells. The transition of ACT from a promising experimental regimen to an established standard of care treatment relies largely on the establishment of safe, efficient, robust and cost-effective cell manufacturing protocols. The manufacture of cellular products under current good manufacturing practices (cGMPs) has a critical role in the process. Herein, we review current manufacturing methods for the large-scale production of clinical-grade TILs, virus-specific and genetically modified CAR or TCR transduced T cells in the context of phase I/II clinical trials as well as the regulatory pathway to get these complex personalized cellular products to the clinic.