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.
Protocol for manufacturing of gene-modified T-memory stem cells (Human Gene Therapy Methods)
… we show the robust clinical-scale production of human peripheral blood T-cells with an early memory phenotype that express a MART-1-specific TCR. By combining selection and stimulation using anti-CD3/CD28 beads for retroviral transduction, followed by expansion in the presence of IL-7 and IL-15, production of a well-defined clinical-scale TCR gene-modified T-cell product could be achieved.
TIL can be rapidly expanded with aAPC to clinical scale generating T cells with similar phenotypic and effector profiles as with PBMC feeders. These data support the clinical application of aAPC to manufacture TIL for the treatment of melanoma.
1. Validation of platelet lysate as alternative to serum in microcarrier-based hMSC expansion (Bioprocess Int) FREE
We investigated whether hPL could be used to expand hMSCs on microcarriers without affecting the cells’ identity and functionality. To that end, we cultured hMSCs on microcarriers and compared their expression of hallmark surface markers and various MSC-defining genes. Thus did we identify hPL as an alternative to FBS for microcarrier-based hMSC cultures.
2. GMP-compliant protocol for manufacturing of T-regs (Cell Transplantation) FREE
Our data provide evidence that functional alloantigen Tregs can be generated under clinical grade compliant conditions. Taking into account that 130 x 106 CD25+ cells can be obtained at large scale from standard leukapheresis, our cell process may give rise to a theoretical final number of 1 x 109 spe-Tregs.
3. Loss of CD34+ cells in hematopoietic products stored in -80C freezer after overnight storage (Transfus Apher Sci)
Our results indicate that the mean loss of post-thaw total and viable CD34+ cells is approximately 20% higher than that observed in standard cryopreservation methods. In addition, fresh viable, post-thaw total and especially post-thaw viable CD34+ cell levels are valuable predictors of both neutrophil and platelet engraftments.
We have optimized the microcarrier-based platform for expansion of MSCs that generated high cell yields in a more efficient and cost-effective manner. This study highlighted the critical parameters in the optimization of MSC production process.
In the present study, we aimed at performing a systemic investigation of the assembling process in perfusion culture for fabricating centimeter-scale macrotissues from cell-laden microcarriers following a bottom-up modular approach.
6. Impact of short-term cold storage on lipoaspirate-derived adipose stem cells (J Plast Reconstr Aesthet Surg)
In summary, we report that human lipoaspirates stored in refrigerator for first 2–3 days after liposuction are still relatively good quality and sterile. This lays the groundwork for potential injection of refrigerated lipoaspirates. However, this result may need to be verified by an in-vivo study using refrigerated lipoaspirates.