Validation of the single-platform ISHAGE protocol for enumeration of CD34+ hematopoietic stem cells in umbilical cord blood in a Brazilian center

Abstract 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.

Celularidade do sangue de cordão umbilical de gestantes hipertensas: estudo caso-controle / Cellularity of the umbilical cord blood of hypertensive pregnant women: a case control study / Celularidad de la sangre de cordón umbilical de gestantes hipertensas: estudio caso-control

OBJETIVO(S): analisar a relação entre celularidade do sangue do cordão umbilical e placentário de gestantes hipertensas e não hipertensas. MÉTODO: estudo caso-controle, com abordagem quantitativa, amostra de 73 gestantes, no período de março a setembro de 2011. Os dados fazem parte do estudo aprovado pelo Comitê de Ética em Pesquisa sob o protocolo de número 126/10. RESULTADOS: foi concluído que 80% das bolsas de sangue de cordão umbilical e placentário coletados de gestantes com hipertensão arterial apresentaram celularidade ≥5 x 108, quantidade adequada no total de células nucleadas, atendendo aos critérios estabelecidos pela Resolução 56. CONCLUSÃO: os resultados contribuíram para a identificação de fatores que possibilitam a obtenção de um quantitativo adequado de células tronco-hematopoiéticas, resultando no aumento do número de transplantes de células tronco-hematopoiéticas no Brasil.

AIM: To analyze the relationship between the cellularity of placental and umbilical cord blood of hypertensive and non-hypertensive pregnant women. METHOD: This is a case-control study that used a quantitative approach. The sample consists of 73 pregnant women and it was conducted from March to September 2011. The data are part of the study approved by the Research Ethics Committee under protocol number 126/10. RESULTS: It was found that 80% of the Umbilical Cord and placental blood bags collected from pregnant women with hypertension presented cellularity ≥5 x 108. This is the appropriate amount in relation to the totality of nucleated cells, given the criteria established by Resolution 56. CONCLUSION: The results contributed to the identification of factors that make it possible to obtain an adequate number of hematopoietic stem cells, resulting in the increased number of hematopoietic stem cell transplantation in Brazil.

OBJETIVO(S): analizar la relación entre celularidad de la sangre del cordón umbilical y placentario de gestantes hipertensas y no hipertensas. MÉTODO: estudio caso-control, con abordaje cuantitativo, muestra de 73 gestantes, en el período de marzo a septiembre de 2011. Los datos hacen parte del estudio aprobado por el Comité de Ética en Investigación bajo el protocolo de número 126/10. RESULTADOS: fue concluido que 80% de las bolsas de sangre de cordón umbilical y placentario colectados de gestantes con hipertensión arterial presentaron celularidad ≥5 x 108, cantidad adecuada en el total de células nucleadas, atendiendo a los criterios establecidos por la Resolución 56. CONCLUSIÓN: los resultados contribuyeron para la identificación de factores que posibilitan la obtención de un cuantitativo adecuado de células madre-hematopoyéticas, resultando en el aumento del número de trasplantes de células madre-hematopoyéticas en Brasil.