Extensive early apoptosis in frozen–thawed CD34-positive stem cells decreases threshold doses for haematological recovery after autologous peripheral blood progenitor cell transplantation

Summary:The number of CD34 þ cells infused into patients at the time of autologous or allogeneic transplantation is a clinically important variable, but the viability of these cells has not been extensively documented. In this study, we analyzed the recovery of viable CD34 þ cells before and after cryopreservation on 79 autologous stem cell products, using a novel flow cytometry assay without red cell lysis. For 70 PBSC harvest samples, the mean viable CD34 þ cell count was 5.98 Â 10 6 /kg (range 0.3-23 Â 10 6 / kg) before freezing and 5.4 Â 10 6 /kg (range 0.2-23 Â 10 6 / kg) after thawing. The median recovery was 93% (range 48-107%), with 90% recovery for NHL (range 48-100%, n ¼ 34), 83% for multiple myeloma (range 56-106%, n ¼ 11), 92.3% for acute leukemia (range 71-100% n ¼ 7) and 94.5% for nonhematological malignancies (range 50-107% n ¼ 18). Similarly, for autologous bone marrows (n ¼ 9) the median recovery of viable CD34 þ cells was 90% (range 68-100%). The recovery of viable CD34 þ cells for adult (n ¼ 51) and pediatric (n ¼ 28) stem cell collections was 91 and 94%, respectively. Further examination of the correlation between the kinetics of hematological recovery and the number of viable progenitor cells infused, particularly at the lower end of the accepted dose range, may be warranted.

Section: Discussionmentioning

confidence: 99%

mentioning

confidence: 99%

Recovery of viable CD34+ cells from cryopreserved hemopoietic progenitor cell products

Sartor

Antonenas

Garvin

et al. 2005

“…from that observed in the five healthy controls (mean 3874.47 s.d., median 38, range [36][37][38][39][40][41][42][43][44]. These data are summarized in Figure 4.…”

Section: Clonogenic Assaymentioning

confidence: 99%

Impaired bone marrow hematopoietic progenitor cell function in rheumatoid arthritis patients candidated to autologous hematopoietic stem cell transplantation

Porta

Caporali

Epis

et al. 2004

Summary:We have evaluated bone marrow morphology, percentage of bone marrow CD34 þ cells, proliferative activity of bone marrow precursors, clonogenic assay (BFU-E and CFU-GM) in short-term bone marrow cultures, and bone marrow cell apoptosis, together with serum TNF-a and IL-6, in 16 chronic, refractory RA patients, as well as in five healthy controls. Of 16 RA patients (68.7%), 11 showed a reduced bone marrow cellularity, while it was normal in all the controls. In RA patients, the median percentage of CD34 þ bone marrow cells, the median percentage of proliferating bone marrow myeloid precursors, and the median number of both BFU-E and CFU-GM colonies were significantly lower than observed in the controls. As far as TNF-a and IL-6 titers is concerned, the latter did not significantly differ from controls' values, while TNF-a titers were significantly lower in healthy controls. Finally, the median apoptotic index of early bone marrow myeloid cells of RA patients was significantly higher compared with controls. These observations may identify the biological risk factors for impaired mobilization and/or engraftment when RA patients are candidates for autologous hematopoietic stem cell grafting.

“…3,4 In spite of these precautions, cryopreservation is associated with variable loss of cell viability and a reduction in the absolute number of viable CD34 þ cells available for reinfusion. [5][6][7] It has been related to the direct effect of low temperature and the formation of ice crystals, 8 or to DMSO only. 9 Infusion of thawed products may induce a wide variety of adverse events usually mild to moderate.…”

Section: Introductionmentioning

confidence: 99%

Recovery, viability and clinical toxicity of thawed and washed haematopoietic progenitor cells: analysis of 952 autologous peripheral blood stem cell transplantations

Foïs

Desmartin

Benhamida

et al. 2007

Cryopreservation and thawing of haematopoietic stem cells are associated with cell loss and infusion-related toxicities. We analysed viability, total nucleated cell (TNC) and CD34 þ cell recovery, and infusion-related toxicities of 952 thawed and washed products. Mean TNC and CD34 þ viable cells recoveries were 55.9718.6 and 98.0736.5%, respectively. Mean cell viability was 68.25718.9%. TNC recovery was correlated with viability but independent of the initial nucleated cell concentration. No difference in TNC recovery or viability was observed according to underlying diseases, except for myeloma, for which these variables were significantly lower (Po0.05). CD34 þ cell recovery was not correlated with viability or CD34 þ initial count and was similar for all diseases. Cryostorage duration was not associated with cell loss. Immediate adverse events occurred in 169 patients (19%) and were moderate (grade I or II) for the majority of patients. Clinical toxicity was associated with a higher infused cell number and the presence of clumps in infused bags. The washing procedure of cell products lead to a low rate of adverse events, but patients transplanted with high cell numbers or bags in which clumps were identified are predisposed to such complications.