Clinical Cell Processing News is a series about new protocols, products and techniques for clinical-grade cell processing and manufacturing. Cell processing devices, cultureware, bioreactors, GMP-grade reagents, cell separation techniques. This series is posted every 2 months.
Review: Toward a scalable and consistent manufacturing process for the production of human MSCs (Cell Gene Ther Insights) FREE
Our findings demonstrate that PIM cryopreservation of HPC(A) products provides recovery of CD34+ cells, CD34+ subpopulations, and CFCs similar to that of DMSO cryopreservation and therefore may have the potential to be used for cryopreservation of peripheral blood stem cells.
This study has demonstrated that HPL, compared with FBS-containing medium, delivers increased growth and comparability across two BM-hMSC donors between monolayer and microcarrier culture, which will have key implications for process transfer during scale-up.
1. Clinical manufacturing of umbilical cord MSC on microcarriers (Stem Cell Int) FREE
… a microcarrier selection out of five microcarrier types was made to achieve a suitable growth surface for the cells. The growth characteristics and metabolite consumption and production were used to compare the cells growth in 12-well plate and spinner flask. The goal to determine relevant process parameters to transfer the expansion process into a stirred tank bioreactor was achieved.
2. Validation of CliniMACS Prodigy for CD34+ cell selection (J Transl Med) FREE
As the clinical GMP-compliant automat CliniMACS Prodigy is being programmed to perform ever more complex sequential manufacturing steps, we developed a CD34+ selection module for comparison with the standard semi-automatic CD34 “normal scale” selection process on CliniMACS Plus, applicable for 600 × 106 target cells out of 60 × 109 total cells. Three split-validation processings with healthy donor G-CSF-mobilized apheresis products were performed; feasibility, time consumption and product quality were assessed.
3. Validation of pulse-activated plasmonic nanobubbles for processing of heterogenoeus human cell grafts (Mol Ther – Methods) FREE
We employ cell-specific on-demand mechanical intracellular impact from laser pulse-activated plasmonic nanobubbles (PNB) to process heterogeneous human cell grafts ex vivo with dual simultaneous functionality, the high cell type specificity, efficacy and processing rate for transfection of target CD3+ cells and elimination of subsets of unwanted CD25+ cells.
4. Optimization of umbilical cord MSC culture for GMP manufacturing (Stem Cell Int) FREE
UC-MSCs were expanded in medium enriched with 2%, 5%, or 10% pooled human platelet lysate (HPL) eliminating the xenogeneic serum components. When the HPL concentrations were compared, media supplemented with 10% HPL had the highest growth rate, smallest cells, and the most viable cells at passage. UC-MSCs grown in 10% HPL had surface marker expression typical of MSCs, high colony forming efficiency, and could undergo trilineage differentiation. The new protocol standardizes manufacturing of UC-MSCs and enables clinical translation.
… we have developed a rapid multiparameter flow cytometric CBU potency assay that enumerates cells expressing high levels of the enzyme aldehyde dehydrogenase (ALDHbr) along with viable CD45+, CD34+ cell content. These measurements are made on a segment that was attached to a cryopreserved CBU. We validated the assay with pre-specified criteria testing Accuracy, Specificity, Repeatability, Intermediate Precision, and Linearity.
PL promotes the rapid expansion of MSCs from CLI and healthy persons. Importantly, MSCs expanded from CLI patients demonstrate the desired angiogenic activity compared with their healthy counterparts. We conclude that autologous MSCs from CLI patients can be sufficiently expanded with PL and be expected to deliver requisite angiogenic effects in vivo.
7. Optimization and modeling of MSC production om microcarrier in stirred bioreactors (Stem Cell Int) FREE
Our established multiregression model makes the rapid definition of the suspension criteria for different working volumes possible and supports the optimization of microcarrier-based, wave-mixed bioreactors used for hASC cultivations. The consideration of further microcarrier types in the regression model would even allow hMSC expansions other than hASCs.
8. DMSO-free cryopreservation of Wharton’s jelly-derived MSC (J Cell Biochem)
In this study, we compared the effect of two different cryoprotectants (DMSO & cocktail solution) on post-thaw cell behavior upon freezing of WJ tissue following two different freezing protocols [Conventional (-1°C/min) & programmed]. The programmed method showed higher cell survival rate compared to conventional method of freezing. Further, cocktail solution showed better cryoprotection than DMSO.”>In this study, we compared the effect of two different cryoprotectants (DMSO & cocktail solution) on post-thaw cell behavior upon freezing of WJ tissue following two different freezing protocols [Conventional (-1°C/min) & programmed]. The programmed method showed higher cell survival rate compared to conventional method of freezing. Further, cocktail solution showed better cryoprotection than DMSO.
9. Validation of hypothermic storage of human pluripotent stem cell-derived cardiomyocytes (Stem Cells TM) FREE
… it was demonstrated that 7 days of cold storage did not affect hPSC-CM ultrastructure, phenotype, or function. This study provides important insights into the cold preservation of PSC-CMs that could be valuable in improving global commercial distribution of hPSC-CMs.