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Establishment of embryonic stem cells secreting human factor VIII for cell‐based treatment of hemophilia A
Author(s) -
KASUDA S.,
KUBO A.,
SAKURAI Y.,
IRION S.,
OHASHI K.,
TATSUMI K.,
NAKAJIMA Y.,
SAITO Y.,
HATAKE K.,
PIPE S. W.,
SHIMA M.,
YOSHIOKA A.
Publication year - 2008
Publication title -
journal of thrombosis and haemostasis
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.947
H-Index - 178
eISSN - 1538-7836
pISSN - 1538-7933
DOI - 10.1111/j.1538-7836.2008.03022.x
Subject(s) - embryonic stem cell , stem cell , microbiology and biotechnology , cancer research , biology , medicine , genetics , gene
Summary. Background: Hemophilia A is an X‐chromosome‐linked recessive bleeding disorder resulting from an F8 gene abnormality. Although various gene therapies have been attempted with the aim of eliminating the need for factor VIII replacement therapy, obstacles to their clinical application remain. Objectives: We evaluated whether embryonic stem (ES) cells with a tetracycline‐inducible system could secrete human FVIII. Methods and results: We found that embryoid bodies (EBs) developed under conditions promoting liver differentiation efficiently secreted human FVIII after doxycycline induction. Moreover, use of a B‐domain variant F8 cDNA (226aa/N6) dramatically enhanced FVIII secretion. Sorting based on green fluorescent protein (GFP)–brachyury (Bry) and c‐kit revealed that GFP–Bry + /c‐kit + cells during EB differentiation with serum contain an endoderm progenitor population. When GFP–Bry + /c‐kit + cells were cultured under the liver cell‐promoting conditions, these cells secreted FVIII more efficiently than other populations tested. Conclusion: Our findings suggest the potential for future development of an effective ES cell‐based approach to treating hemophilia A.