11^th Annual Conference on Stem Cell and Regenerative Medicine October 15-16, 2018 Amsterdam, Netherlands

Saba Rezaei-Lotfi has completed her MPhil in Faculty of Dentistry, Department of Life Sciences. Her thesis provided novel experimental evidence regarding the regulation of neurogenesis by microRNA signaling. She is currently a PhD student at the Faculty of Medicine and Health. Her research project focuses on investigating the role of Catenin-β1 in determining fate in neural differentiation

Statement of the Problem: Neural self-organization refers to spontaneous evolution of global order from local interactions between neural progenitor cells during neurogenesis. However, the molecular basis for programming various aspects of neural self-organization including sub-lineage fate choice of the progenitor cells remains ambiguous. Here we report the role of Catenin-β1 in encoding the programmable dimension of neural self-organization based on subcellular localization of this protein. Method: We used human neural progenitors with neuronal and glial differentiation capacities in the current study. Prior to neural induction, small molecule inhibitors were applied to program the subcellular localization of Catenin-β1. Subsequent to programming, the proliferation rate of the progenitors was assessed using live imaging phase contrast microscopy. Differentiation of the programmed cells into neuronal and glial elements was then investigated using a combination of immuno-histochemical staining and real-time PCR. Findings: The application of IWR-1 resulted in depletion of cytoplasmic free Catenin-β1 by mobilization into junctional N-cadherin. The IWR-1+ cells also demonstrated increased nuclear notch-1 and down regulation of YAP-1. The application of ML-141, on the other hand, mobilized the junctional Catenin-β1 into the cytoplasmic pool and subsequently into the nucleus, increased activity of free Catenin-β1 lead to the depletion of antagonistic Notch-1 in these cells. While Catenin-β1+/Notch-1- neural progenitors adopted a neuronal fate, the Catenin-β1-/Notch-1+ cells mainly differentiated into glial elements. Conclusion: Subcellular localization of Catenin-β1 is a major determinant of neurogenic fate bias. Localization of this protein to functional complexes anticipates a glial fate bias and mobilization into the cytoplasmic pool instructs a neuronal differentiation bias. These findings may be utilized to program the progenitor cells in guided tissue engineering.

Malaria is a lethal endemic disease, causing over million deaths annually. The destruction of erythrocytes by P. falciparum and thrombocytopenia by P. vivax causes anemia, major co morbidity. Adverse effects are more prominent in children and pregnant women. The available treatments for anemia are transfusion of freshly collected RBC, erythropoietin injection, blood substitutes and dietary supplementation of hematinic. The major challenge is the unavailability of safe screened blood in the developing and under developed countries, for immediate transfusion to the patient. Cord blood contains CD34+ cells, fetal hemoglobin 60%-80% (oxygen carrying capacity is 60% more than the adult hemoglobin), higher platelet and WBC content, hypo-antigenic with altered metabolic profile, anti-malarial effect. The cytokines and growth factors of the cord blood have stimulatory effect on bone marrow. The institutional ethical committee approved the study, for which 94 units of cord blood obtained from healthy consenting mothers via LUCS . The collected volume of cord blood/placenta varied 52 ml to 143ml mean packed cell volume 48.9±4.1 SD and mean hemoglobin concentration 16.4 g%±1.6 g% SD. Cord blood was screened and transfused to 39 consenting randomly selected patients suffering from confirmed malaria (varying between the age of 8years to 72 years) with hemoglobin less than 8 g% (pre-transfusion hemoglobin varied between 5.4 g/dl to 7.4 g/dl), according to standard blood transfusion protocol (screened, cross matched between donor and recipient). Rise in hemoglobin after 72 hours of transfusion was (0.5 g/dl-1.6 g/dl). On 7th day, statistically significant (p<0.003) rise in hemoglobin was observed, also peripheral CD34+ cells increased. No detectable rise in glucose, serum creatinine, urea or bilirubin was observed. No immunological or non-immunological adverse reaction was observed. Cord blood is an under-utilized, potentially advantageous substitute for adult blood transfusion in cases of severe anemia.

Suvodip Chakrabarty has completed his MBBS and MPhil in Regenerative Medicine.

Among the most debilitating disabilities of the present day, rheumatoid arthritis has a significant position affecting almost 0.8% of Western population and about 7 million in Indian population (2010). Anemia is a very common comorbidity of rheumatoid arthritis. Like anemia of chronic illness, it is caused by defective iron metabolism, coexistent intestinal parasites, steroids and NSAIDs, Methotrexate and Sulphasalazine. If the hemoglobin is 8 g/dl or less, blood transfusion is recommended along with replacement for iron deficiency, but in case of rheumatoid arthritis anemia does not effectively utilize iron to form hemoglobin. Because of higher proportion of fetal hemoglobin (about 70%), cytokines and growth factors, being hypo antigenic nature, we hereby propose that human umbilical cord blood has properties comparable (and at times superior) to adult blood transfusion. 83 units (45 ml-137 ml, mean packed cell volume 44.3±2.1 SD, mean hemoglobin concentration 16.1 g/dl±1.5 g/dl SD) of placental umbilical cord whole blood were transfused to 32 informed consenting patients with advanced rheumatoid arthritis, who had hemoglobin of 8 g/dl or less. The patients received two to six units of freshly collected placental umbilical cord blood without encountering any recognizable/visible clinical, immunological or non-immunological reactions. Peripheral blood hematopoietic stem cell (CD34+) estimation revealed a rise from the pre-transfusion base level (0.092%) by 7 to 10 days, varying from 2.03 to 22%. Mean rise in hemoglobin with 2 units cord blood transfusions varied from 0.5 gm/dl to 1.5 gm/dl after 72 hours and about 0.2 gm/dl to 0.9 gm/dl after 7-10 days from transfusion. In our study, we conclude the feasibility of transfusion of whole cord blood in patients with anemia in rheumatoid arthritis, with no discernible immunological reactions so far in last 19 years.