BMI1 enables extensive expansion of functional erythroblasts from human peripheral blood mononuclear cells

Mol Ther. 2021 May 5;29(5):1918-1932. doi: 10.1016/j.ymthe.2021.01.022. Epub 2021 Jan 21.

Abstract

Transfusion of red blood cells (RBCs) from ABO-matched but genetically unrelated donors is commonly used for treating anemia and acute blood loss. Increasing demand and insufficient supply for donor RBCs, especially those of universal blood types or free of known and unknown pathogens, has called for ex vivo generation of functional RBCs by large-scale cell culture. However, generating physiological numbers of transfusable cultured RBCs (cRBCs) ex vivo remains challenging, due to our inability to either extensively expand primary RBC precursors (erythroblasts) or achieve efficient enucleation once erythroblasts have been expanded and induced to differentiation and maturation. Here, we report that ectopic expression of the human BMI1 gene confers extensive expansion of human erythroblasts, which can be derived readily from adult peripheral blood mononuclear cells of either healthy donors or sickle cell patients. These extensively expanded erythroblasts (E3s) are able to proliferate exponentially (>1 trillion-fold in 2 months) in a defined culture medium. Expanded E3 cells are karyotypically normal and capable of terminal maturation with approximately 50% enucleation. Additionally, E3-derived cRBCs can circulate in a mouse model following transfusion similar to primary human RBCs. Therefore, we provide a facile approach of generating physiological numbers of human functional erythroblasts ex vivo.

Keywords: BMI1; RBC; enucleation; erythroblasts; gene editing; sickle cell disease; transfusion.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

  • Adult
  • Animals
  • Cell Culture Techniques
  • Cell Differentiation
  • Cell Proliferation
  • Cells, Cultured
  • Erythroblasts / cytology*
  • Erythrocyte Transfusion / methods*
  • Erythrocytes / cytology*
  • Fetal Blood
  • Humans
  • Leukocytes, Mononuclear / cytology*
  • Mice
  • Models, Animal
  • Polycomb Repressive Complex 1 / genetics*

Substances

  • BMI1 protein, human
  • Polycomb Repressive Complex 1