Expression of coagulation factor IX in human CD34 + hematopoietic stem cells by adeno-associated virus 2.
- Author:
Yan CHEN
1
;
Fang-ping CHEN
;
Jian-qiang PENG
;
Xiao-bing WU
;
Guang-ping WANG
;
Zai-fu JIAN
;
Hong-ya XIN
;
Xin-hua WU
Author Information
- Publication Type:Journal Article
- MeSH: Antigens, CD34; Cell Differentiation; Cell Proliferation; Cells, Cultured; Dependovirus; genetics; Factor IX; genetics; metabolism; Fetal Blood; cytology; Gene Expression; Genetic Vectors; Hematopoietic Stem Cells; cytology; immunology; metabolism; Humans; RNA, Messenger; genetics; Transfection
- From: Chinese Journal of Hematology 2005;26(9):529-533
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVETo investigate the expression of human coagulation factor IX (hFIX) gene in human umbilical cord blood (CB) CD34+ cells which was transfected with recombinant adeno-associated virus 2 (rAAV-2).
METHODThe CD34+ cells were transfected with rAAV-2/hFIX and cultured for 21 days for inducing differentiation into myeloid, erythroid and megakaryocytes, respectively. The expression of hFIX was studied at mRNA, protein and biological activity levels. The cytotoxicity of rAAV-2 to CD34+ cells was evaluated by cell proliferation, cell vitality, CD antigen expressions and CFU yields.
RESULTSThe hFIX mRNA was expressed in the cultured cells which was verified by RT-PCR and DNA sequencing. An elevated level of hFIX expression and biological activities were detected with a maximum amount of 14.10 ng/10(6) cell x 24 h. During the period of 21 day culture, the cell vitality, cell proliferation, CD antigen expression and CFU yields between the transfected and un-transfected groups had no difference(P > 0.05).
CONCLUSIONThe human CB CD34+ cells are able to produce functional hFIX after transduction by rAAV-2/hFIX. The cell proliferation and differentiation capacities of the host CD34+ cells were not affected by rAAV-2.
