Reproducible establishment of hemopoietic supportive stromal cell lines from murine bone marrow.
Stromal cell lines, designated MS-1, -2, -3, -4, -5, -6, and -7 were established by irradiating the adherent cells in long-term bone marrow cultures with 900-rad x-rays. Two of the cell lines, MS-1 and MS-5, have the capacity to support the growth of hemopoietic stem cells (spleen colony-forming cells and granulocyte-macrophage colony-forming cells) for greater than 2 months in vitro. These two cell lines were alkaline phosphatase-, peroxidase-, and factor VIII-negative and positive for periodic acid-Schiff and nonspecific esterase. Extracellular matrix proteins such as fibronectin, laminin, and collagen type I were produced by these two cell lines. Neither MS-1 cell- nor MS-5 cell-conditioned medium supported the growth of hemopoietic stem cells, and hemopoietic stem cells were found preferentially to be under and on MS-1 and MS-5 layers rather than in suspension. Close contact with the MS-1 cell layer or the MS-5 cell layer appears to be essential in maintaining hemopoiesis in vitro. Conditioned media from MS-1 cells and MS-5 cells stimulated granulocyte colony formation from murine bone marrow cells in semisolid culture.
Itoh
K
K
Department of Biology, Faculty of Science, Niigata University, Japan.
Tezuka
H
H
Sakoda
H
H
Konno
M
M
Nagata
K
K
Uchiyama
T
T
Uchino
H
H
Mori
K J
KJ
D000818:Animals; D001853:Bone Marrow; D002448:Cell Adhesion; D002460:Cell Line; D003114:Colony-Forming Units Assay; D003115:Colony-Stimulating Factors / Q000096:biosynthesis; D005109:Extracellular Matrix / Q000032:analysis / Q000378:metabolism / Q000502:physiology*; D005260:Female; D006410:Hematopoiesis; D006412:Hematopoietic Stem Cells / Q000166:cytology / Q000502:physiology* / Q000528:radiation effects; D006651:Histocytochemistry; D008297:Male; D051379:Mice; D008809:Mice, Inbred C3H; D013997:Time Factors
D003115:Colony-Stimulating Factors