Spectrum of hemoglobin disorders in southern Odisha, India: a hospital based study.

BACKGROUND: Hemoglobin disorders are the leading health concern in the world including India. There is a paucity of literature on the spectrum of hemoglobin disorders in southern districts of Odisha state. This study was undertaken to elucidate the occurrence of different hemoglobin disorders in a tertiary health care facility of Odisha state, India. METHODS: The study cases were suspected patients of all age groups advised for screening of different hemoglobin disorders. Hemoglobin disorders were screened by sickling slide test and high-performance liquid chromatography (HPLC) using the Variant-II hemoglobin testing system as per the manufacturer's guidelines. RESULTS: Over 2 years, 2332 blood samples (including 1102 pediatric and 1230 adult cases) were investigated, out of which, 1380 (59.2%) of cases had abnormal hemoglobin disorders. The most common was sickle cell disorders (48.67%, 1135/2332) followed by ß-thalassemia (11.32%, 264/2332). Some rare variants were detected as hemoglobin D-Punjab, hemoglobin E, hemoglobin Lepore, hereditary persistence of fetal hemoglobin, hemoglobin with high P2 window, hemoglobin with high P3 window etc, Among the cases with abnormal hemoglobin disorders, 744 (53.9%), 545 (39.5%) and, 91 (6.6%) cases were found to have the heterozygous, homozygous and, double heterozygous state. Of the 188 ante-natal cases screened, 31.4% of cases had abnormal hemoglobin variants with sickle cell disorders being the most prevalent one. CONCLUSION: Along with the high occurrence of sickle cell disorders in the study area, some other rare hemoglobin disorders are also prevalent which calls for a large community-based cohort study.

Association of CD34+ and CD90+ Stem Cells of Cord Blood with Neonatal Factors: A Cross-sectional Study.

OBJECTIVE: To characterize the primitive stem cell content of cord blood with regard to neonatal parameters. METHODS: In this cross-sectional study, CD34+ and CD90+ cells content were enumerated by flow-cytometry method. Their associations with various neonatal parameters like birth weight, gender, gestational age and mode of delivery were analyzed by univariate analysis. Multivariable linear regression model was then developed to further explain the effect of neonatal factors on these primitive cell counts. RESULTS: From a total of 106 recruited subjects, gender of the neonate did not have any influence on the expression of these proteins (CD34 and CD90) of cord blood stem cells or progenitors. Multi variable linear regression analysis using CD34+ and CD90+ cell counts as dependent variables revealed that birth weight and the mode of delivery were significant predictors of these cell counts. CONCLUSIONS: The present study suggests that birth weight and mode of delivery of the neonates influences cord blood stem cell yield.

Cryoprotective Effect of Disaccharides on Cord Blood Stem Cells with Minimal Use of DMSO.

Umbilical cord blood (UCB) is an extremely attractive source of stem cells for the treatment of various benign and malignant hematological and non-hematological disorders. To facilitate the preservation of these stem cells, 10 % dimethylsulfoxide (DMSO) is widely used as cryoprotectant in cord blood banks. But it is found to be toxic at this concentration with the result of serious side effects in recipients after infusion of DMSO-cryopreserved cells. Evaluation of viability and functionality of cryopreserved hematopoietic stem cells is needed with either inclusion of nontoxic additives alone or with reduced DMSO concentration. We assessed the post thawing viability of UCB stem cells in the freezing medium containing disaccharides (sucrose or trehalose) alone and in combination with reduced amount i.e. 2 % DMSO by trypan blue staining. The functionally active progenitor cells content of the optimized media was then evaluated and compared with 5% DMSO by a colony forming unit assay using methylcellulose based media. The freezing solution containing 0.2 M trehalose with 2 % DMSO came out to be superior in the evaluation of viability and generation of hematopoietic colonies of erythroid and myeloid lineage than 5 % DMSO alone. While the percentage of viability was lower than 2 % DMSO, as observed in the medium containing 0.2 M trehalose or sucrose alone, with poor outcome of generation of myeloid lineage based colonies. Our study results suggest that trehalose (0.2M) with the inclusion of reduced concentration of DMSO(2%) can better replace 5%DMSO rather than complete removal of DMSO from the freezing medium.

In-vitro Behavior of Human Umbilical Cord Blood Stem Cells Towards Serum Based Minimal Cytokine Growth Conditions.

We tried here to optimize the proliferation of both Hematopoietic and Mesenchymal stem cells of Umbilical Cord blood in minimal cytokine growth condition. Failing to get good results of expansion of non-adherent Hematopoietic Total Nucleated Cells and adherent Fibroblastic Mesenchymal Stem Cells derived from 10-12 ml of collected Cord blood, we designed the further experimental study by increasing the volume of Cord blood sample up to 65-70 ml. We harvested the non-adherent as well as adherent fraction separately derived from the primary culture of Umbilical Cord blood stem cells under the influence of growth promoting Cytokines or Growth Factors. The proliferation study was conducted by taking different combinations of two hematopoietic growth stimulatory Cytokines like stem cell factor (SCF) and Fms like tyrosine kinase-3Ligand (Flt3L) at concentrations (10 ng/ml, 100 ng/ml) while we preferred Mesenchymal specific growth factor i.e. basic Fibroblast growth factor (FGF-ß) at its 10 ng/ml concentration for adherent cells to get optimal results. The Hematopoietic and Fibroblast Colony forming abilities of the expanded stem cells were performed through Colony Forming Unit assay. Culture Medium containing cytokine combination like SCF 100 ng/ml with Flt3L 10 ng/ml was found to be optimal for the proliferation of hematopoietic stem cells. But the number of hematopoietic colonies like Erythroid colonies generated were less in case of media supplemented with SCF & Flt3L while more number of Myeloid colonies were observed in Growth factor supplemented media in comparison to the control one. The FGF-ß supplemented media successfully enhanced the proliferation of Mesenchymal Stem Cells and exhibited its efficient Fibroblast colony forming ability. Our experimental study supports the minimal utilization of cytokines for haematopoietic and mesenchymal stem cell proliferation which may help in future safe Cord blood stem cell infusion.