Characterization of Cultured Human Erythrocytes Derived from BMI-1 Transduced Erythroblasts: Towards an Alternate Blood Supply Restricted; Files Only

Abstract

There is a need for an alternate blood supply to counteract donor shortages, provide a reliable source for atypical transfusions in cases of alloimmunization, and rare blood groups and for reagent cells in the blood bank.[1, 2] Erythrocytes (RBCs) produced ex vivo may provide an inexhaustible alternative or complimentary source of blood products for those patients without compatible donors, while eliminating the risk of infection and transfusion reactions.[3] Scalable ex vivo RBC production for transfusion purposes requires a substrate that is self-renewing and has high expansion capacity. Bmi-1 overexpression was first shown to increase the self-renewal potential of murine erythroblasts and we previously published studies employing Bmi-1 overexpression in human erythroblasts to produce cultures of extensively expanded erythroblasts (E3s).[4, 5]  Here we highlight our in vitro culture technique of erythroid maturation from E3s in a manner that is reproducible and demonstrate that E3s can be differentiated into mature erythrocytes (cRBCs). Further we extensively characterized the resultant cRBCs derived from E3s. We demonstrate that cRBCs are in the reticulocyte stage of maturation and that these cultured reticulocytes (cRetics) are similar in structure and function to native reticulocytes (nRetics) found in blood circulation. Our findings support the use of Bmi-1 transduced E3s as a viable substrate for to be used for the ex-vivo production of RBCs as an alternate blood source for transfusion especially for those patients who may not otherwise have a suitable source for transfusion. 

Table of Contents

Table of Contents

Chapter 1         Introduction                                                                             Page 4

Chapter 2         Materials and Methods                                                          Page 6

Chapter 3         Results                                                                                      Page 10

Chapter 4         Discussion                                                                                Page 13

Chapter 5         Figures and tables                                                                  Page 15

                          References                                                                                Page 24      

About this Master's Thesis

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School	Laney Graduate School
Department	Clinical Research
Degree	M.S.
Submission	Master's Thesis
Language	English
Research Field	Health Sciences, Medicine and Surgery Biology, Cell
Palavra-chave	Cultured red cells
Committee Chair / Thesis Advisor	Amita Manatunga, Emory University
Committee Members	John Roback, Emory University David Jaye, Emory University

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