Methods for production of platelets from pluripotent stem cells and compositions thereof
AssigneeAstellas Institute for Regenerative Medicine
InventorsQiang Feng, Shi-Jiang Lu, Robert P. Lanza
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Abstract
Methods for production of platelets from pluripotent stem cells, such as human embryonic stem cells (hESCs) and induced pluripotent stem cells (iPSCs) are provided. These methods may be performed without forming embryoid bodies or clusters of pluripotent stem cells, and may be performed without the use of stromal inducer cells. Additionally, the yield and/or purity can be greater than has been reported for prior methods of producing platelets from pluripotent stem cells. Also provided are compositions and pharmaceutical preparations comprising platelets, preferably produced from pluripotent stem cells.
Claims14
A method for producing β2 microglobulin-deficient platelets from megakaryocytes comprising:
- contacting a non-adherent feeder-free culture of β2 microglobulin-deficient megakaryocytes with hematopoietic expansion medium and
- (1) thrombopoietin (TPO) or a TPO agonist, stem cell factor (SCF), interleukin (IL)-6, and IL-9;
- or
- (2) TPO or a TPO agonist, SCF, and IL-11;
- or
- (3) TPO or a TPO agonist
- to cause the formation in culture of pro-platelets that release β2 microglobulin-deficient platelets, at least 60% of which are positive for CD41a and CD42b expression.
A method of producing platelets, comprising:
- (a) culturing pluripotent stem cells engineered to be β2 microglobulin deficient in a feeder-free culture comprising TPO or a TPO agonist to form-megakaryocytes;
- and
- (b) contacting a non-adherent feeder-free culture of the megakaryocytes with hematopoietic expansion medium and
- (1) thrombopoietin (TPO) or a TPO agonist, stem cell factor (SCF), interleukin (IL)-6, and IL-9, or
- (2) TPO or a TPO agonist, SCF, and IL-11, or
- (3) TPO or a TPO agonist
- to cause the formation in culture of pro-platelets that release platelets, at least 60% of which are positive for CD41a and CD42b expression.
A method of producing platelets, comprising
- contacting a non-adherent feeder-free culture of β2 microglobulin knockout megakaryocytes with hematopoietic expansion medium and
- (1) thrombopoietin (TPO) or a TPO agonist, stem cell factor (SCF), interleukin (IL)-6, and IL-9, or
- (2) TPO or a TPO agonist, SCF, and IL-11, or
- (3) TPO or a TPO agonist
- to cause the formation in culture of pro-platelets that release platelets, at least 60% of which are positive for CD41a and CD42b expression.
A method of producing platelets, comprising
- contacting a non-adherent feeder-free culture of HLA-A-deficient megakaryocytes with hematopoietic expansion medium and
- (1) thrombopoietin (TPO) or a TPO agonist, stem cell factor (SCF), interleukin (IL)-6, and IL-9, or
- (2) TPO or a TPO agonist, SCF, and IL-11, or
- (3) TPO or a TPO agonist
- to cause the formation in culture of pro-platelets that release platelets, at least 60% of which are positive for CD41a and CD42b expression.
Dependencies are inferred from the claim text.