ABSTRACT
BACKGROUND AND OBJECTIVES: Our post-thaw cell recovery rates differed substantially in interlaboratory comparisons of identical samples, potentially due to different temperatures during cell staining. MATERIALS AND METHODS: Viable CD34(+) cells and leucocyte (WBC) subtypes were quantified by multiparameter single-platform flow cytometry in leucapheresis products collected from 30 adult lymphoma and myeloma patients, and from 10 paediatric patients. After thawing, cells were prepared for analysis within 30 min between thawing and acquisition, at either 4°C or at room temperature. RESULTS: For cell products cryopreserved in conventional freezing medium (10% final DMSO), viable cell recovery was clearly lower after staining at 4°C than at RT. Of all WBC subtypes analysed, CD4(+) T cells showed the lowest median recovery of 4% (4°C) vs. 25% (RT), followed by CD3, CD34 and CD8 cells. The recovery was highest for CD3γδ cells with 44% (4°C) vs. 71% (RT). In the 10 samples cryopreserved in synthetic freezing medium (5% final DMSO), median recovery rates were 89% for viable CD34 (both at 4°C and RT) and 79% (4°C) vs 68% (RT) for WBC. CONCLUSIONS: The post-thaw environment and, potentially, the cryoprotectant impact the outcome of cell enumeration, and results from the analysis tube may not be representative of the cells infused into a patient.
Subject(s)
Leukocytes/cytology , Adult , Antigens, CD34/metabolism , Flow Cytometry , Freezing , Humans , Leukocytes/metabolism , Multiple Myeloma , Staining and Labeling , TemperatureSubject(s)
Anemia, Refractory/genetics , Anemia, Sideroblastic/genetics , Calreticulin/genetics , Mutation/genetics , Thrombocytosis/genetics , Adult , Aged , Anemia, Refractory/diagnosis , Anemia, Sideroblastic/diagnosis , Cohort Studies , Female , Humans , Male , Middle Aged , Prognosis , Thrombocytosis/diagnosis , Young AdultSubject(s)
Anemia, Refractory/genetics , Janus Kinase 2/genetics , Myelodysplastic Syndromes/blood , Myelodysplastic Syndromes/genetics , Polymorphism, Single Nucleotide/genetics , Receptors, Thrombopoietin/genetics , Aged , Aged, 80 and over , Cohort Studies , DNA Mutational Analysis , Female , Humans , Male , Middle Aged , Platelet Count , RegistriesSubject(s)
Chromosomes, Human, Pair 5 , Genetic Markers , Janus Kinase 2/genetics , Myeloproliferative Disorders/genetics , Myeloproliferative Disorders/pathology , Adult , Aged , Aged, 80 and over , Bone Marrow/pathology , Cell Division , Chromosome Deletion , Female , Humans , Male , Middle Aged , Myeloproliferative Disorders/classification , Point MutationABSTRACT
AIMS: One-hundred and eighty-eight cases of human mammary carcinoma were examined immunohistochemically for their expression of Ki67, p34cdc2 and c-erbB-2. DNA image cytometry was performed to evaluate DNA ploidy, Auer type, S-phase fraction (SPF), 5c exceeding rate (5cER) and 2c deviation index (2cDI). METHODS AND RESULTS: One-hundred and sixty-eight cases were invasive ductal carcinomas, 20 were of invasive lobular type. Routinely assessed oestrogen and progesterone receptor scores were available. The results were analysed statistically in comparison to tumour type, histopathological grade, lymph node status, menopausal status, patient age and overall survival. Ki67 (P < 0.002) and c-erbB-2 (P < 0.0001) correlated well with overall survival (P < 0.0008) and grade (P < 0.038) but not with lymph node status and tumour type. p34cdc2 showed a trend towards a positive correlation with Ki67 (P < 0.058) and a significant negative correlation with receptor status (P < 0.008) but with none of the other parameters examined. CONCLUSIONS: No association between the DNA measured parameters (Auer type, SPF, 5cER and 2cDI) and survival was found. Our results suggest that c-erbB-2 and Ki67 are parameters which might, in combination with receptor status, help to define subgroups with different outcomes.
