Changes in hematopoiesis-supporting ability of C3H10T1/2 mouse embryo fibroblasts during differentiation.
To investigate the functional change of stromal cells along with differentiation, we used a differentiation-inducible mouse embryo fibroblast cell line, C3H10T1/2 (10T1/2). Stably determined preadipocyte and myoblast cell lines were established after a brief exposure of 10T1/2 cells to 5-azacytidine. These cell lines terminally differentiated into adipocytes and myotubes, respectively, under appropriate conditions. The hematopoiesis-supporting ability of each 10T1/2-derived cell line was examined by coculture with FACS-sorted murine hematopoietic stem cells (Thy-1lo c-kit+ Lin-). The number of granulocyte-macrophage progenitors (CFU-GM) was slightly reduced after 7 days of culture with parent 10T1/2 fibroblasts, whereas a marked increase in CFU-GM number was observed when the cells were cultured on preadipocytes. Mature adipocytes and myogenically determined cell lines, on the other hand, did not support CFU-GM growth. Further, Northern analysis showed that the preadipocyte cell line acquired the ability to produce a significant amount of stem cell factor (SCF), interleukin-6 (IL-6), leukemia inhibitory factor, and macrophage colony-stimulating factor mRNAs in response to IL-1 or lipopolysaccharide stimulation. Terminal adipocytic differentiation resulted in reduced ability to express these cytokine mRNAs. Similarly, highest IL-6 activity was detected in the supernatant of preadipocyte culture, whereas adipocytes did not secrete IL-6 even after IL-1 stimulation. Interestingly, hematopoiesis-nonsupporting myoblasts and myotubes also expressed abundant SCF mRNA, suggesting that SCF, per se, may not be sufficient for stem cell growth and survival. The 10T1/2-derived cell lines could provide a valuable tool to aid in the analysis of stromal cell development and the search for novel stromal factors.
Nishikawa
M
M
Department of Hematology-Oncology, University of Tokyo, Japan.
Ozawa
K
K
Tojo
A
A
Yoshikubo
T
T
Okano
A
A
Tani
K
K
Ikebuchi
K
K
Nakauchi
H
H
Asano
S
S
D000273:Adipose Tissue / Q000166:cytology; D000818:Animals; D001483:Base Sequence; D002454:Cell Differentiation; D002460:Cell Line; D016207:Cytokines / Q000096:biosynthesis / Q000235:genetics; D004622:Embryo, Mammalian; D005109:Extracellular Matrix / Q000502:physiology; D005260:Female; D005347:Fibroblasts; D006410:Hematopoiesis; D016298:Hematopoietic Cell Growth Factors / Q000235:genetics; D051379:Mice; D008809:Mice, Inbred C3H; D008810:Mice, Inbred C57BL; D008969:Molecular Sequence Data; D009132:Muscles / Q000166:cytology; D011247:Pregnancy; D012333:RNA, Messenger / Q000032:analysis; D019089:Stem Cell Factor; D017154:Stromal Cells / Q000502:physiology*
D016207:Cytokines; D016298:Hematopoietic Cell Growth Factors; D012333:RNA, Messenger; D019089:Stem Cell Factor