- Helene Lapillonne1,
- Ladan Kobari2,
- Christelle Mazurier3,
- Philippe Tropel4,
- Marie-Catherine Giarratana5,
- Isabelle Zanella-Cleon6,
- Laurent Kiger7,
- Marie Wattenhofer-Donze8,
- Helene Puccio8,
- Nicolas Hebert9,
- Alain Francina10,
- Georges Andreu11,
- Stephane Viville8 and
- Luc Douay12,*
- 1 INSERM UMR_S938 and UPMC Paris6 and AP-HP, France;
- 2 INSERM UMR_S938 and UMPC Paris, France;
- 3 INSERM UMR_S938 and EFS Ile de France, France;
- 4 IGBMC and INSERM/UEVE UMR-861, France;
- 5 INSERM UMR_S938 and UPMC Paris, France;
- 6 IBCP, France;
- 7 INSERM U473 and Hopital Bicetre, France;
- 8 IGBMC, France;
- 9 INSERM UMR_S938 and UPMC Paris 6, France;
- 10 Unite' de Pathologie Moleculaire du Globule Rouge, Hopital Edouard Herriot, France;
- 11 INTS, France;
- 12 INSERM UMR_S938 and UPMC Paris 6 and AP-HP and EFS, France
- ↵* Corresponding author; email:
Background. Ex vivo manufacture of Red Blood Cells (RBC) from stem cells is a possible means to ensure an adequate and safe supply of blood cell products. Advances in somatic cell reprogramming to human induced pluripotent stem cells (hiPSCs) have opened the door to generating specific cells for cell therapy. HiPSCs represent a potentially unlimited source of stem cells for erythroid generation for transfusion medicine.
Design and Methods. We characterized the erythroid differentiation and maturation of hiPSCs cell lines obtained from human fetal (IMR90) and adult fibroblasts (FD-136) compared to hESC line (H1). Our protocol comprises two steps: (1) differentiation of hiPSCs by formation of embryoid bodies with indispensable conditioning in the presence of cytokines and human plasma to obtain early erythroid commitment and (2) differentiation/maturation to the stage of cRBC in the presence of cytokines. The protocol dispenses with major constraints such as obligatory passage through a hematopoietic progenitor, coculture on a cellular stroma and use of proteins of animal origin.
Results. We report for the first time the complete differentiation of hiPSCs into definitive erythropoïesis capable of maturation up to enucleated RBCs containing fetal hemoglobin in a functional tetrameric form.
Conclusions. RBCs generated from hiPSCs pave the way for future development of allogeneic transfusion products. This could be done at least using the banking of a very limited number of red cell phenotype combinations enabling to transfuse safely a great number of immunized patients.
- Received February 8, 2010.
- Accepted May 17, 2010.
- Copyright © 2010, Ferrata Storti Foundation