Characterization of murine stromal cell clones established from bone marrow and spleen.
Stromal cell lines were established by irradiating adherent layers of bone marrow and spleen cells in Dexter-type long-term culture with X-rays. Some of these cell lines support myelopoiesis and/or B lymphopoiesis in vitro. Furthermore, the characteristics of these stromal cell lines were studied. Cytokine activity was detected in the conditioned media from all hematopoietic-supportive and non-supportive stromal cells. Quantitative reverse transcriptase-polymerase chain reaction analysis revealed that the mRNAs of macrophage colony-stimulating factor and stem cell factor, but not that of Interleukin-3, were detectable in all the hematopoietic-supportive and non-supportive stromal cell lines. The transcripts of granulocyte colony-stimulating factor, interleukin-6, interleukin-7, and leukemia inhibitory factor were expressed in a wide variety of cell lines. Most stromal cell lines synthesized mRNA of c-mpl, the ligand of which stimulates megakaryopoiesis and thrombopoiesis. These observations indicate that the pattern of mRNA expression of cytokines is not correlated with the hematopoietic-supportive ability of stromal cell lines. There was a significant difference in the efficiency of adhesion of lineage marker-negative bone marrow cells to fibroblasts and stromal cell lines. This appears to be correlated with the hematopoietic-supportive ability of the stromal cell lines.
Tsuji
T
T
Life Science Research Laboratory, JT Inc, Yokohama, Japan.
Ogasawara
H
H
Aoki
Y
Y
Tsurumaki
Y
Y
Kodama
H
H
D016475:3T3 Cells; D000818:Animals; D001402:B-Lymphocytes / Q000166:cytology; D001483:Base Sequence; D015415:Biomarkers; D001854:Bone Marrow Cells; D002448:Cell Adhesion; D002460:Cell Line; D019070:Cell Lineage; D018920:Coculture Techniques; D003238:Connective Tissue / Q000502:physiology; D003239:Connective Tissue Cells; D005347:Fibroblasts / Q000378:metabolism; D015870:Gene Expression; D006410:Hematopoiesis / Q000502:physiology; D016298:Hematopoietic Cell Growth Factors / Q000096:biosynthesis / Q000235:genetics; D006412:Hematopoietic Stem Cells / Q000528:radiation effects; D051379:Mice; D008807:Mice, Inbred BALB C; D008809:Mice, Inbred C3H; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction; D012333:RNA, Messenger / Q000096:biosynthesis / Q000235:genetics; D013154:Spleen / Q000166:cytology*
D015415:Biomarkers; D016298:Hematopoietic Cell Growth Factors; D012333:RNA, Messenger