Comparative quantification of umbilical cord blood CD34+ and CD34+ bright cells using the ProCount™-BD and ISHAGE protocols

Abstract

The total number of CD34+ cells is the most relevant clinical parameter when selecting human umbilical cord blood (HUCB) for transplantation. The objective of the present study was to compare the two most commonly used CD34+ cell quantification methods (ISHAGE protocol and ProCount - BD) and analyze the CD34+ bright cells whose 7-amino actinomycin D (7AAD) analysis suggests are apoptotic or dead cells. Twenty-six HUCB samples obtained at the Placental Blood Program of New York Blood Center were evaluated. The absolute numbers of CD34+ cells evaluated by the ISHAGE (with exclusion of 7AAD+ cells) and ProCount (with exclusion of CD34+ bright cells) were determined. Using the ISHAGE protocol we found 35.6 +/- 19.4 CD34+ cells/microL and with the ProCount method we found 36.6 +/- 23.2 CD34+ cells/microL. With the ProCount method, CD34+ bright cell counts were 9.3 +/- 8.2 cells/microL. CD34+ bright and regular cells were individually analyzed by the ISHAGE protocol. Only about 1.8% of the bright CD34+ cells are alive, whereas a small part (19.0%) is undergoing apoptosis and most of them (79.2%) are dead cells. Our study showed that the two methods produced similar results and that 7AAD is important to exclude CD34 bright cells. These results will be of value to assist in the correct counting of CD34+ cells and to choose the best HUCB unit for transplantation, i.e., the unit with the greatest number of potentially viable stem cells for the reconstitution of bone marrow. This increases the likelihood of success of the transplant and, therefore, the survival of the patient.

Patient age and number of apheresis days may predict development of secondary myelodysplastic syndrome and acute myelogenous leukemia after high‐dose chemotherapy and autologous stem cell transplantation for lymphoma

“…A total of 3× the total blood volume was processed using an apheresis system (COBE Spectra, Terumo BCT). Stem cells were quantified using ISHAGE protocol . The goal was to collect 5 × 10 6 CD34+ cells/kg in six or fewer apheresis sessions, with a minimum acceptable cell dose of 2 × 10 6 CD34+ cells/kg for transplantation.…”

Colheita de medula óssea em cães: modelo para obtenção da fração total de células mononucleares

“…O transplante de MO para terapia apresenta algumas limitações para sua utilização, como a obtenção da fração total de CM em quantidade suficiente, que exige múltiplas aspirações da MO em ambiente cirúrgico (GUNSILIUS, 2001). Outro aspecto limitante é o tempo médio para se obter efeito terapêutico depois de transplantadas, cerca de duas a quatro semanas, isto é, para a geração das novas células sangüíneas (PRANKE et al, 2006) ou para o fornecimento dos elementos celulares necessários durante o processo regenerativo como a quimiotaxia e o efeito parácrino em um novo microambiente (LEVENBERG et al 2003).…”

Predicting overall viability of cord blood harvests

“…As birth weight increased, so did volume, total nucleated cell count, CD34+ cell count, and colony‐forming unit–granulocyte‐macrophage count 12 . However, the total number of CD34+ cells decreases with gestational age 12,13 …”