Research Project
Grant-in-Aid for Scientific Research (C)
The study purpose is to establish the gene transfer system and its evaluation method for a growth failure of one of any lineage of hematopoietic progenitors caused by a single gene abnormality We focused on the glycogen storage disease type 1 b associated with congenital neutropenia caused by mutations in glucose 6-translocase (G6PT)gene. First, G6PT gene expression vector(pCS-CDF-ChG6PT-PRE) was developed. Then CD34-positive cells derived from healthy umbilical cord blood were purified and G6PT gene (or GFP gene) was transferred to the cells by the lentivirus vector. After transfection, methylcellulose colony assay was carried out to evaluate the gene transfer efficiency. Then bone marrow cells from a patient with glycogen storage disease type Ib were collected.G6PT or GFP gene was introduced to the purified CD34-positive cells using the vectors. Methylcellulose colony assay was performed to evaluate the forming ability of G-CFU,GM-CSF,M-CSF and E-BFU colonies. And the cells from each colony were collected and calculated the transduction efficiency by PCR.As a result, gene transfer efficiency was as low as about 10 percent, and gene transfer effect was not significantly improved the forming ability of any type colonies in methylcellulose medium. To find the optimal conditions, this study is now under the way.
All 2008 2007 2006 2005 Other
All Journal Article (24 results) (of which Peer Reviewed: 12 results) Presentation (16 results)
Pediatrics 121
Pages: e892-9
Pediatric Research Feb;63(2)
Pages: 182-5
Human Genetics 121(2)
Pages: 223-232
Clinical Endocrinology 67
Pages: 212-217
Journal of Human Genetics 52(12)
Pages: 1040-1043
Hum Genet. 121(2)
Pages: 223-32
Clin Endocrinol. 67
J Hum Genet. 52(12)
Pages: 1040-3
Int J Immunogenet 33(1)
Pages: 7-10
Diabetic Med 23(10)
Pages: 1145-50
Am J Neuroradiol 27(9)
Pages: 1924-6
Diabetic Med. 23(10)
Am J Neuroradiol. 27(9)
Pediatr Transplant 9(1)
Pages: 101-103
J Mol Med 83(4)
Pages: 316-320
J Inherit Metab Dis 28(5)
Pages: 681-687
Exp Hematol 33
Pages: 1215-1221
Pediatr Res. (in press)
Pediatrics. (in press)
