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Artigo em Inglês | MEDLINE | ID: mdl-33358615


BACKGROUND: This study aims to validate the single-platform method for enumeration of CD34+ cells, by comparing the performance of two different commercial kits, as well as to evaluate the efficiency of the AccuriTM C6 cytometer in providing direct counts of absolute cell numbers. METHOD: We evaluated 20 samples from umbilical cord blood (UCB), comparing the two different methodologies for enumeration of CD34+ cells: single and dual-platform. For the assessment of the single-platform, Procount and SCE kits were used, both of which use fluorescent beads as a counting reference to obtain absolute CD34+ cells numbers. Moreover, after the acquisition of samples in flow cytometer AccuriTM C6, following the protocol established for each kit, the number of CD34+ cells was recalculated, considering the cell count provided by the AccuriTM C6. MAIN RESULTS: In our analysis, the results showed a strong correlation between the number of CD34+ cells/µL (r2=0.77) when comparing the SCE kit and the current dual-platform method. On the other hand, the comparison between Procount kit and dual-platform results showed a moderate correlation for the number of CD34+/µL cells (r2=0.64). CONCLUSION: Our results showed that the AccuriTM C6 flow cytometer can be used safely, applying both the dual and single platform analysis strategy. Considering the ISHAGE protocol-based single-platform approach, as the most appropriate methodology for CD34+ cells enumeration, our results demonstrated that the SCE kit has great potential for national standardization of UCB samples analysis methodology.

Cytotherapy ; 21(3): 327-340, 2019 03.
Artigo em Inglês | MEDLINE | ID: mdl-30685216


Clinical trials of adoptively transferred CD19 chimeric antigen receptor (CAR) T cells have delivered unprecedented responses in patients with relapsed refractory B-cell malignancy. These results have prompted Food and Drug Administration (FDA) approval of two CAR T-cell products in this high-risk patient population. The widening range of indications for CAR T-cell therapy and increasing patient numbers present a significant logistical challenge to manufacturers aiming for reproducible delivery systems for high-quality clinical CAR T-cell products. This review discusses current and novel CAR T-cell processing methodologies and the quality control systems needed to meet the increasing clinical demand for these exciting new therapies.

Imunoterapia Adotiva/métodos , Instalações Industriais e de Manufatura/normas , Neoplasias/terapia , Controle de Qualidade , Receptores de Antígenos Quiméricos , Antígenos CD19/imunologia , Remoção de Componentes Sanguíneos/métodos , Sobrevivência Celular , Criopreservação/métodos , Endotoxinas/análise , Humanos , Imunoterapia Adotiva/efeitos adversos , Ativação Linfocitária , Mycoplasma , Linfócitos T/imunologia , Transdução Genética/métodos