Immortalization of bone marrow-derived human mesenchymal stem cells by removable simian virus 40T antigen gene: analysis of the ability to support expansion of cord blood hematopoietic progenitor cells.
Human marrow-derived mesenchymal stem cells (MSC), which have the potential to differentiate into mesenchymal tissues, such as bone, cartilage, adipose and bone marrow stroma, were transduced with a retroviral vector carrying the simian virus 40 large T antigen, hygromycin-resistant gene and herpes simplex virus thymidine kinase gene, that can be excised by Cre/loxP site-specific recombination. This resulted in establishment of an MSC cell line, HMSC-1, which retained original surface characteristics and differentiation potential, and exhibited a higher proliferative capacity than parental cells. HMSC-1 expressed mRNAs of BMP-4, Jagged-1, and SCF that are known to promote hematopoiesis. Human CB CD34+ hematopoietic progenitor cells (HPC) cultured on a layer of HMSC-1 cells showed high expansion of CD34+CD38- immature HPC, capable of reconstituting human hematopoiesis in non-obese diabetic/severe combined immunodeficient disease (NOD/SCID) mice. This cell line may be of value for developing strategies for ex vivo expansion of human HPC.
Nishioka
Keisuke
K
Division of Hematology-Oncology, Department of Medicine, Hyogo College of Medicine, Hyogo 663-8501, Japan. fuji-y@hyo.med.ac.jp
Fujimori
Yoshihiro
Y
Hashimoto-Tamaoki
Tomoko
T
Kai
Shunro
S
Qiu
Huiying
H
Kobayashi
Naoya
N
Tanaka
Noriaki
N
Westerman
Karen A
KA
Leboulch
Philippe
P
Hara
Hiroshi
H
D036541:ADP-ribosyl Cyclase / Q000096:biosynthesis; D051997:ADP-ribosyl Cyclase 1; D000818:Animals; D015703:Antigens, CD / Q000096:biosynthesis; D018952:Antigens, CD34 / Q000096:biosynthesis; D000952:Antigens, Polyomavirus Transforming / Q000235:genetics*; D015153:Blotting, Western; D001854:Bone Marrow Cells / Q000166:cytology*; D018929:Cell Culture Techniques / Q000379:methods; D002454:Cell Differentiation; D002934:Cinnamates / Q000494:pharmacology; D018920:Coculture Techniques; D003114:Colony-Forming Units Assay; D005312:Fetal Blood / Q000166:cytology*; D005434:Flow Cytometry; D006412:Hematopoietic Stem Cells / Q000166:cytology*; D006801:Humans; D006921:Hygromycin B / Q000031:analogs & derivatives* / Q000494:pharmacology; D008562:Membrane Glycoproteins; D008648:Mesoderm / Q000166:cytology*; D051379:Mice; D016513:Mice, SCID; D008957:Models, Genetic; D010641:Phenotype; D012333:RNA, Messenger / Q000378:metabolism; D011995:Recombination, Genetic; D012190:Retroviridae / Q000235:genetics; D020133:Reverse Transcriptase Polymerase Chain Reaction; D013234:Stem Cells / Q000166:cytology*; D013997:Time Factors
D015703:Antigens, CD; D018952:Antigens, CD34; D000952:Antigens, Polyomavirus Transforming; D002934:Cinnamates; D008562:Membrane Glycoproteins; D012333:RNA, Messenger; D006921:Hygromycin B; C026273:hygromycin A; D036541:ADP-ribosyl Cyclase; C498154:CD38 protein, human; C498155:Cd38 protein, mouse; D051997:ADP-ribosyl Cyclase 1