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

Azel Zine has his expertise in the study of development and regeneration of the peripheral auditory system. He has made seminal contributions to our understanding of the Notch signaling and HES gene roles in the development of auditory hair cells in mammals. His scientific contributions have all been well thought out and comprehensive, published in some of highly peer reviewed journals. His work with applying the degradation resistant JNK inhibitor (D-JNK-1) to the inner ear as an otoprotective drug has also been notable and the results reported in that initial work has led to clinical trials in Europe that are in progress under the direction of Auris Medical-Biotechnology Company. His current research is in the field of stem cells biology.

Deafness is a major public health issue and the most common sensory deficit in humans. Approximately 360 million people have disabling hearing losses, a number likely to grow due to increasing noise pollution, ototoxic drugs and aging. Most forms of deafness are progressive and neurodegenerative disorders involving the loss of sensory hair cells and their associated primary auditory neurons. These sensory cells are not replaced and hearing loss is permanent. A stem cell-based therapy could in principle offers reasonable expectations for the potential treatment of inner ear disorders through the replacement of lost or damaged sensory cells. Initial advances in the differentiation of murine ESCs/iPSCs into hair cell and neuron-like cells have paved the way for similar progresses with human pluripotent stem cells. In this study, we used monolayer cultures, exposure to otic-inducing agents, Notch signaling modulation, and cell type marker expression to obtain characterized human otic/placodal progenitors from human induced Pluripotent Stem Cells (hiPSCs). Then, we explored the engraftment ability of in vitro generated human otic progenitor cells in experimental model of sensorineural hearing loss. The results from our study indicate that hiPSC-derived otic/placodal progenitor cells survived up to one month after transplantation, migrated and integrated into the endogenous cochlear epithelium of in vivo ototoxic model of hearing loss. Once within the microenvironment of the ototoxic damaged cochlea, some of the injected human otic progenitor’s up regulate a subset of initial inner ear sensory cell type markers. Information’s provided by the experiments of this study would bring the possibility of using a stem cell-based cell therapy as a potential option for deafness closer to becoming reality and pave the way for clinical trials in human.

Niranjan Bhattacharya has completed his MBBS and MD in Obstetrics and Gynecology. He has pursued his Master’s degree in General Surgery and a DSc in Developmental Immunology. His principal specializations are cell and tissue therapy. He has many publications in international and national journals on cord blood and regenerative medicine. He is the co-editor of five books on the subject published by Springer. He is currently working as a Chair, Professor and Head of the Department, Regenerative Medicine and Translational Science and Director General, first Public Cord Blood Bank in India, Calcutta School of Tropical Medicine, Kolkata.

The term blood substitute is actually a misnomer because only a part of the total functions of the blood is replaced by any available so-called substitute i.e. oxygen delivery and volume expansion only. Therefore, a more accurate term should be red cell substitute. Cord blood, because of its rich mix of fetal and adult hemoglobin, high platelet and WBC counts and a plasma filled with cytokine and growth factors, as well as its hypo antigenic nature and altered metabolic profile, has all the potential of a real and safe alternative to adult blood during emergencies due to any etiology of blood loss and anemia. Our experience of 192 units of cord blood transfusion in patients with beta thalassemia with severe anemia (hemoglobin concentration varying from 3.5 to 6 g/dl with mean hemoglobin 4.67 g/dl) proved to be extremely effective in 84 patients as an emergency substitute of adult RBC transfusion (male: female ratio 1:1, age varying from 6 months to 38 years). In the present series, the collection of the blood varied from 57 ml-136 ml mean 84 ml±7.2 ml SD, median 87 ml, mean packed cell volume 45±3.1 SD, mean hemoglobin concentration 16.4 g/dl±1.6 g/dl SD. After collection the blood was immediately preserved in the refrigerator and transfused within 72 hours of collection from the consenting mother undergoing lower uterine cesarean section. We did not encounter a single case of immunological or non immunological reaction till date. We suggest that the medical fraternity use this precious gift of nature, which is free from infection, hypo antigenic with an altered metabolic profile, filled with growth factor and cytokine filled plasma with potential higher oxygen carrying capacity than for adult blood, as an emergency source of blood for the management of transfusion-dependent beta thalassemia.

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.

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 143 ml 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.