چکیده (انگلیسی):

Hemophilia A is an X-linked rescessive bleeding disorder that results from F8 gene aberrations.
Previously, we established embryonic stem (ES) cells (tet-226aa/N6-Ainv18) that secrete
human factor VIII (hFVIII) by introducing the human F8 gene in mouse Ainv18 ES cells. Here,
we explored the potential of cell transplantation therapy for hemophilia A using the ES cells.
Transplant tet-226aa/N6-Ainv18 ES cells were injected into the spleens of severe combined
immunodeficiency (SCID) mice, carbon tetrachloride (CCl4)-pretreated wild-type mice, and
CCl4-pretreated hemophilia A mice. F8 expression was induced by doxycycline in drinking
water, and hFVIII-antigen production was assessed in all cell transplantation experiments.
Injecting the ES cells into SCID mice resulted in an enhanced expression of the hFVIII antigen;
however, teratoma generation was confirmed in the spleen. Transplantation of ES cells into
wild-type mice after CCl4-induced liver injury facilitated survival and engraftment of transplanted
cells without teratoma formation, resulting in hFVIII production in the plasma.
Although CCl4 was lethal to most hemophilia A mice, therapeutic levels of FVIII activity, as well
as the hFVIII antigen, were detected in surviving hemophilia A mice after cell transplantation.
Immunolocalization results for hFVIII suggested that transplanted ES cells might be engrafted at
http://dx.doi.org/10.1016/j.hemonc.2016.04.002
1658-3876/
2016 King Faisal Specialist Hospital & Research Centre. Published by Elsevier Ltd.
This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/).
Abbreviations: Bry, brachyury; CCl4, carbon tetrachloride; DAPI, 40,6-diamidino-2-phenylindole; Dox, doxycycline; EB, embryoid body; ES,
embryonic stem; FVIII, factor VIII; FVIII:Ag, factor VIII antigen; FVIII:C, factor VIII activity; FIX, factor IX; GFP, green fluorescent protein;
H&E, hematoxylin and eosin; hFVIII, human factor VIII; iPS, induced pluripotent stem; LSEC, liver sinusoidal endothelial cell; PCR, polymerase
chain reaction; SCID, severe combined immunodeficiency; tet, tetracycline; VWF, von Willebrand factor; WT, wild type
* Corresponding author at: Department of Pediatrics, Matsubara Tokushukai Hospital, 7-13-26 Amami-higashi, Matsubara, Osaka 580-0032,
Japan.
E-mail address: ysakurai-th@umin.ac.jp (Y. Sakurai).