Cell separation by counterflow centrifugal elutriation has been described for the preparation of monocytes for vaccine applications, but its use in other current good manufacturing practice (cGMP) operations has been limited. In this study, growth factor-mobilized peripheral blood progenitor cell products were collected from healthy donors and processed by elutriation using a commercial cell washing device. Fractions were collected for each product as per the manufacturer's instructions or using a modified protocol developed in our laboratory. Each fraction was analyzed for cell count, viability, and blood cell differential. Our data demonstrate that, using standard elutriation procedures, >99% of red blood cells and platelets were removed from apheresis products with high recoveries of total white blood cells and enrichment of CD34؉ cells in two of five fractions. With modification of the basic protocol, we were able to collect all of the CD34؉ cells in a single fraction. The CD34-enriched fractions were formulated, labeled with a ferromagnetic antibody to CD34, washed using the Elutra device, and transferred directly to a magnetic bead selection device for further purification. CD34؉ cell purities from the column were extremely high (98.7 ؎ 0.9%), and yields were typical for the device (55.7 ؎ 12.3%). The processes were highly automated and closed from receipt of the apheresis product through formulation of target-enriched cell fractions. Thus, elutriation is a feasible method for the initial manipulations associated with primary blood cell therapy products and supports cGMP and current good tissue practice-compliant cell processing. STEM CELLS TRANSLATIONAL MEDICINE 2012;1:422-429
Background: The achievement of optimal number of CD34 + umbilical cord stem cells is essential for successful umbilical cord stem cell transplantation. So the aim of this study was to assess the potential effect of both maternal and neonatal factors on the umbilical cord blood CD34 + cell count. Methods:The study was done on umbilical cord blood samples obtained from 20 mothers during labor.Their ages ranged from 22 to 34 years and were subjected to history taking, physical examination of the baby and assessment of the CD34 + cells count in umbilical cord blood.Results: Number of previous live births and weight of the baby had a significant effect on CD34 + cells count while the sex of the baby, delivery route, maternal age and gestation period had no significant effect on CD34 + cells count.Conclusions: Umbilical cord blood-derived CD34 + cell count is better with good weight and first babies and decreased with subsequent babies.
Background: Nocturnal enuresis (NE) is a very common chronic pediatric problem with bad psychological consequences. Methods: Forty primary monosymptomatic nocturnal enuresis (MNE) children and 20 healthy controls were recruited in the study and subjected to history taking, neurological and urological examinations, and psychological assessment using the Arabic-translated and validated version of child behavior checklist, sleep architecture studying through onenight polysomnography (PSG), and vasopressin levels determination both diurnal and nocturnal. Results: Enuretic children had positive family history of NE in 42.5%, inverted vasopressin circadian rhythm in 52.5% and PSG changes in the form of increased N3 deep sleep % of total sleep time (TST), sleep stage transition index (SSTI), periodic limb movement index (PLMI), and snore index. Enuretic children PSG showed decreased deep sleep latency, N1% of TST, N2% of TST, and REM % of TST. The child behavior checklist showed higher anxious depressed symptoms, social problem, attention problems, and internalizing problems in enuretic children than control subjects. Conclusions: MNE is a heterogeneous disorder with multiple factors interplay in its pathogenesis. So, the management must be tailored patient by patient according to the dominating etiology.
Background Phototherapy (PT) is the most often utilized technique for treating and preventing severe hyperbilirubinemia in the term and preterm newborns. PT's proven benefit is that it decreases the requirement for exchange transfusions. To investigate the effect of PT on allergic response mediators in neonates with hyperbilirubinemia treated by PT, eosinophil counts and tumor necrosis factor alfa levels have been assessed. Methods This cross-sectional study included 100 full-term infants with indirect hyperbilirubinemia in the first two weeks of life who were indicated for PT. They were investigated by tumor necrosis factor α and eosinophil counts before and 72 h after starting PT. The used tests were paired with Student’s t-test and Pearson coefficient. Results Relative and absolute eosinophil counts and tumor necrosis factor alfa were significantly higher after PT than before (p < 0.001). There was a significant positive correlation between total serum bilirubin and both tumor necrosis factor alfa and eosinophil % (r = 0.442 and r = 0.362, respectively, P < 0.001) before PT. There was a significant positive correlation between total serum bilirubin and both eosinophil count and eosinophil % (r = 0.281and r = 0.339), respectively (P < 0.001) after PT. There was a significant positive correlation between both tumor necrosis factor alfa and eosinophil % after PT (r = 0.545, P < 0.001). Conclusions Serum tumor necrosis factor-alpha and eosinophilic count increased after treatment of neonatal hyperbilirubinemia by PT, which indicates an allergic response to PT in neonates.
Background: Allogeneic hematopoietic stem cell transplantation (HSCT) is the only commonly available curative treatment for people with thalassemia major (TM).This work aimed to study the impact of using low dose versus high dose anti thymocyte globulin (ATG (based conditioning regimen on the results of peripheral blood stem cell transplantation in children with βeta- TM. Methods: This comparative study was established on 40 patients with βeta-thalassemia major (Pesaro class II and/or III) who subjected to (HSCT) from donor who is related. Our patients received the same myeloablative regimen. They were then categorized according to dose of ATG into two groups: group I (Busulfan/Cyclophosphamide/Low dose ATG-30 mg) and group II (Busulfan/Cyclophosphamide/High dose ATG-110 mg). Results: There was no association between both groups according to the clinical features, transplant related complications in both groups. There was a higher risk of mucositis and infections in high dose group. (P = 0.024 & P = 0.046; respectively). Patients in low dose group achieved faster neutrophil engraftment with median value of (14 days). No difference was found in the incidence of post-transplant viral reactivation, graft versus host disease in both groups. Conclusions: Pesaro II, III thalassemia patients can safely receive hematopoietic stem cell transplantation (HSCT) with low dose ATG-30 mg regimen and attain the same result.
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