Concise Review:

Stem Cell-Based Approaches to Red Blood Cell Production for Transfusion

Presenter ： Zhangyu 2015.11.4

Presenter ： Yu Zhang

2015.11.4

www.who.int/worldblooddonorday/en/

1. Introduction

Anna Rita Migliaccio, et al. Blood Reviews 26 (2012) 81–95.

Luc DouayInstitut National de la Transfusion Sanguine, Paris, Lutetia Parisorum, Île-de-France, France

Article: Human induced pluripotent stem cells can reach complete terminal maturation: in vivo and in vitro evidence in the erythropoietic differentiation model. •Haematologica 06/2012; · 6.42 Impact Factor

Article: Banking of pluripotent adult stem cells as an unlimited source for red blood cell production: potential applications for alloimmunized patients and rare blood challenges. •Transfusion medicine reviews 03/2011; 25(3):206-16. · 3.61 Impact Factor

Article: Human fetal liver: an in vitro model of erythropoiesis. •Stem cells international. 01/2011; 2011:405429.

Article: Red blood cell generation from human induced pluripotent stem cells: perspectives for transfusion medicine. •Haematologica 10/2010; 95(10):1651-9. · 6.42 Impact Factor

Article: Unimpaired terminal erythroid differentiation and preserved enucleation capacity in myelodysplastic 5q(del) clones: a single cell study. Haematologica 10/2009; 95(3):398-405. · 6.42 Impact Factor

Article: Ex vivo expansion does not alter the capacity of umbilical cord blood CD34+ cells to generate functional T lymphocytes and dendritic cells. •Stem Cells 10/2006; 24(9):2150-7. · 7.78 Impact Factor

Article: Ex vivo generation of fully mature human red blood cells from hematopoietic stem cells. •Nature Biotechnology 02/2005; 23(1):69-74. · 23.27 Impact Factor

Article: Human erythroid cells produced ex vivo at large scale differentiate into red blood cells in vivo. •Nature Biotechnology 06/2002; 20(5):467-72. · 23.27 Impact Factor

2. Hematopoiesis & Erythropoiesis hematopoiesis

Fig.1 　 The developmental process of hematopoiesis in early embryo of mouse.

Mao Bin, et al. Chinese Journal of Cell Biology 2012, 34(11): 1067–1079

2. Hematopoiesis & Erythropoiesis Erythropoiesis

Fig.2 　 A current model of erythropoiesis.

Siddharth Shah, et al. Stem cells translational medicine,2014,3:346-355.

3. Ex vivo Production of Human RBCs Source of stem cells for the production process

3. Ex vivo Production of Human RBCs Summary of current protocols for in vitro RBC generation.

Vimal K. Singh, et al. Cell and developmental bilolgy. 2014, (1),1-18.

4. The biological principles of ex-vivo erythroblasts (EBs) expansion

Cell size & Surface makers

Ann Zeunera, et al. Stem Cells. 2012 August ; 30(8): 1587–1596.Anna Rita Migliaccio, et al. Blood Reviews 26 (2012) 81–95.

4. The biological principles of ex-vivo EB expansion Hemoglobin (Hb) 1、组成

2、种类胚型血红蛋白 ε 、 ζ，胎儿型血红蛋白 γ, 成体型血红蛋白 δ 、 β

正常成人红细胞中有 3种血红蛋白： 成人血红蛋白： HbA （ α2β2) 96%-98%

成人次要血红蛋白 :HbA2 （ α2δ2) 1%-3.1%

胎儿血红蛋白 :HbF （ α2γ2) 2%以下胚胎血红蛋白 : HbG1( ζ2ε2) HbG2 (α2ε2 ) <5 周 HbP( ζ2γ2) 3周 -12周 Fig.A. Fig.3 The oxyhemoglobin dissociation

curve

Figure 3. Hemoglobin analyses and functionality of erythroid cells generated from hiPSCSCD.

4. The biological principles of ex-vivo EB expansion Hb analysis & Oxygen transport capacity & Deformability measurements

Ladan Kobari ,et al. Hematopoiesis & Hematopoietic stem cells 2012;97(12):1795-1803.

4. The biological principles of ex-vivo EB expansion Erythrocyte-specific gene expression

Honglian Jin, et al. BioMed Research International,2014,1-9

4. The biological principles of ex-vivo EB expansion Enucleation Mechanisms

Fig.5 Key events in cytokinesis and enucleation.

Fig.6 In vivo maturation of cRBCs in the NOD/SCID mouse model.

In vivo maturation of cRBC in the NOD/SCID mouse.

4. The biological principles of ex-vivo EB expansion

Marie-Catherine Giarratana, et al. Blood, 2011, 118: 5071-5079

Introduction-Aim

10 million RBCs (the equivalent of 2 ml of blood) were generated ex vivo under good manufacturing practice (GMP) conditions from CD34pos cells obtained by apheresis from an adult volunteer.

The survival in vivo of these expanded autologous RBC labeled with 51Cr(the only method accepted by the U.S. Food and Drug Administration) was comparable to that of native RBC.

Siddharth Shah, et al. Stem cells translational medicine,2014,3:346-355.

Ann Zeunera, et al. Stem Cells. 2012 August ; 30(8): 1587–1596.

6. Prospective time table for clinic application of ex-vivo expanded erythrocytes.

Anna Rita Migliaccio, et al. Blood Reviews 26 (2012) 81–95.

Discussion

Thank you！