Murine serum obtained from bone marrow-transplanted mice promotes the proliferation of hematopoietic stem cells by co-culture with MS-5 murine stromal cells.
To examine whether serum obtained from bone marrow-transplanted mice can selectively expand hematopoietic stem cells (HSCs) among whole bone marrow cells in vitro, whole bone marrow cells were cultured with or without MS-5 murine stromal cells in the presence of serum obtained from transplanted mice on day 3 (day 3 serum) or serum from normal mice for 7 days. When whole bone marrow cells and MS-5 cells were co-cultured in day 3 serum for 7 days, the c-kit-positive, Sca-1-positive, lineage marker-negative cells (KSL cells) expanded approximately 25 times; however, when they were co-cultured in normal serum for 7 days, the KSL cells expanded approximately 1.3 times. Direct contact between the whole bone marrow cells and MS-5 cells was essential for expansion of KSL cells in the co-culture, and it upregulated the expression of some cytokines in MS-5. Above all, the day 3 serum specifically upregulated the expression of SCF, SDF-1 alpha, G-CSF, IL-11 and IL-6 in MS-5. The level of testosterone in the day 3 serum was higher than normal serum and the addition of the testosterone in the culture expanded the KSL cells among whole bone marrow cells on MS-5 cells and also upregulated the expression of SDF-1 alpha, IL-11 and IL-6 in MS-5. These data indicates that the serum of bone marrow-transplanted mice contains a factor(s) that induced changes in the expression levels of various cytokines in MS-5 stromal cells and enabled the MS-5 cells to expand the KSL cells among whole bone marrow cells.
Nakayama
Akemi
A
Faculty of Graduate School of Science and Technology, Department of Cell Science, Niigata University, Niigata, Japan.
Matsui
Haruna
H
Fukushima
Teruaki
T
Ichikawa
Hiroshi
H
Yamada
Kensuke
K
Amao
Takuji
T
Hosono
Masamichi
M
Sugimoto
Kenkichi
K
D000818:Animals; D000950:Antigens, Ly / Q000378:metabolism; D015415:Biomarkers / Q000378:metabolism; D001854:Bone Marrow Cells / Q000166:cytology / Q000378:metabolism; D016026:Bone Marrow Transplantation; D049109:Cell Proliferation; D002478:Cells, Cultured; D018920:Coculture Techniques; D003470:Culture Media; D006412:Hematopoietic Stem Cells / Q000166:cytology* / Q000378:metabolism; D008297:Male; D008565:Membrane Proteins / Q000378:metabolism; D051379:Mice; D019009:Proto-Oncogene Proteins c-kit / Q000378:metabolism; D044967:Serum / Q000378:metabolism*; D013154:Spleen / Q000166:cytology; D017154:Stromal Cells / Q000166:cytology / Q000378:metabolism; D013739:Testosterone / Q000097:blood; D015854:Up-Regulation
D000950:Antigens, Ly; D015415:Biomarkers; D003470:Culture Media; C074650:Ly6a protein, mouse; D008565:Membrane Proteins; D013739:Testosterone; D019009:Proto-Oncogene Proteins c-kit