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.
1. Optimization of bone marrow cells with platelet rich plasma preparation for local applications (J Appl Biomater Funct Mater)
For an optimized stable gel clot, human BMC and PRP should be combined with 10% to 20% fibrinogen (9 mg/mL to 18 mg/mL) and 5% to 20% thrombin (25 I.E. to 100 I.E.). Both freshly prepared and stored cells for 1 to 7 days had a stable consistence over 28 days at 37°C. Different platelet concentrations did not influence gel clot formation. The ratio of living cells did not decrease significantly over the observation period of 5 days in the live-dead staining.
2. Preparation of neural stem cell suspensions for intra-cerebral transplantation (Cell Transplant) FREE
NSCs were suspended in five different vehicles: Phosphate Buffered Saline (PBS), Dulbecco’s Modified Eagle Medium (DMEM), artificial Cerebral Spinal Fluid (aCSF), Hypothermosol and Pluronic. Suspension accuracy, consistency, and cell settling were determined for different cell volume fractions in addition to cell viability, cell membrane damage and clumping. Maintenance of cells in suspension was evaluated while these were stored for 8 hours on ice, at room temperature, or physiological normothermia.
3. Comparison of different cryosolutions for cryopreservation of cord blood (Stem Cells Int) FREE
The low concentration of DMSO with the addition of trehalose might improve the cryopreservation outcome.
Ejections at slower flow rates resulted in a lower percentage of the cell dose being delivered, and apoptosis measurements of samples ejected at 10 µl/minute were significantly higher than control samples. Immunophenotyping also revealed significant downregulation of CD105 expression in samples ejected at 10 µl/minute.
5. Optimization of NK cell isolation and clinical production (J Immunother)
This study aimed to optimize clinical grade production of a cytokine-activated NK-cell product. NK cells were isolated either by double depletion (CD3, CD19) or by sequential depletion and enrichment (CD3, CD56) via CliniMACS from leukapheresis material and cultured in vitro with interleukin (IL)-2 or IL-15.
6. Comparison of commercially available clinical grade human platelet lysate for MSC expansion (Scand J Clin Lab Invest)
MSCs from bone marrow (BMSCs) and adipose tissue-derived stromal cells (ASCs) obtained from three donors were culture expanded in three different commercially available hPL fulfilling good manufacturing practice criteria for clinical use. BMSCs and ASCs cultured in Minimum Essential Medium Eagle-alpha supplemented with 5% PLT-Max (Mill Creek), Stemulate™ PL-S and Stemulate™ PL-SP (COOK General Biotechnology) were compared to standard culture conditions with 10% fetal bovine serum (FBS). Cell morphology, proliferation, phenotype, genomic stability, and differentiation potential were analyzed.
We have optimized methods for generating pancreatic cancer-specific TILs that can be used for adoptive cellular therapy of patients with pancreatic cancer.
Cryopreservation of a CD34+-selected product can be safely used in a combined transplant with UCB and does not affect engraftment time.