Reprogramming T cells and hematopoietic cells
Methods and compositions relating to the production of induced pluripotent stem cells (iPS cells) are disclosed. For example, induced pluripotent stem cells may be generated from CD34 + hematopoietic cells, such as human CD34 + blood progenitor cells, or T cells. Various iPS cell lines are also provided. In certain embodiments, the invention provides novel induced pluripotent stem cells with a genome comprising genetic rearrangement of T cell receptors.
1. A method for producing human induced pluripotent stem cells from CD34 + hematopoietic progenitor cells comprising the steps of:
(a) obtaining a cell population comprising human CD34 + hematopoietic progenitor cells, wherein the CD34 + hematopoietic progenitor cells are from a subject whose cells have not been mobilized; and
(b) producing iPS cells from the CD34 + hematopoietic progenitor cells of the cell population through the introduction of nucleic acids encoding reprogramming factors selected from Oct4, Sox2, cMyc, Klf4, Nanog and Lin28, the nucleic acids being comprised in one or more episomal vectors, and culturing the cells under defined, feeder-free conditions to provide a human iPS cell population that is essentially free of integrated, exogenous viral elements.
2. The method of claim 1 , wherein the source of the cell population is a blood sample, blood components, bone marrow, lymph node, fetal liver, or umbilical cord.
3. The method of claim 2 , wherein the source of the cell population is a blood sample of from about 1 to about 5 ml.
4. The method of claim 3 , wherein the source of the cell population is a blood sample of about 3 ml.
5. The method of claim 1 , wherein the cell population has been cryopreserved.
6. The method of claim 1 , wherein the cell population CD34 + hematopoietic progenitor cells is cultured in vitro with one or more cytokines to expand the T cell or CD34 + hematopoietic progenitor cells population therein.
7. The method of claim 6 , wherein the one or more cytokines comprises at least one of SCF, Flt3L, or IL-3.
8. The method of claim 1 , wherein the reprogramming factors are encoded on one or more expression cassettes that comprise at least one polycistronic transcript unit.
9. The method of claim 8 , wherein the polycistronic transcript unit comprises at least two reprogramming genes.
10. The method of claim 8 , wherein the polycistronic transcript unit comprises a reprogramming gene and a selectable or screenable marker.
11. The method of claim 1 , wherein producing iPS cells from the CD34 + hematopoietic progenitor cells of the population further comprises selecting the iPS cells for one or more characteristics of embryonic stem cells.
12. The method of claim 11 , wherein the characteristic is an adherent property, an undifferentiated morphology, an embryonic stem cell-specific marker or pluripotency.
13. The method of claim 12 , wherein the characteristic is an adherent property.
14. The method of claim 12 , wherein the characteristic is an undifferentiated morphology.
15. The method of claim 1 , wherein the method further comprises differentiating the iPS cells to a differentiated cell.
16. The method of claim 15 , wherein the differentiated cell comprises a cardiomyocyte, a hematopoietic cell, a neuron, a fibroblast or an epidermal cell.