Neural cell adhesion molecule contributes to hemopoiesis-supporting capacity of stromal cell lines.
To clarify mechanisms underlying cell-to-cell interactions between hemopoietic stem cells (HSCs) and stromal cells, we established a stromal cell line (FMS/PA6-P) from day-16 fetal bone marrow (BM) adherent cells using an anti-PA6 monoclonal antibody (mAb) specific for BM stromal cells. Importantly, this FMS/PA6-P cell line, showing homogenous fibroblastic morphology, is absent from hematolymphoid and endothelial lineage markers and maintains a high level of expression of PA6 molecule, recognized by the anti-PA6 mAb, for approximately 20 passages. Further, the cell line expressing a high level of PA6 molecule has a better hemopoiesis-supporting capacity in vitro than other stromal cell lines such as PA6 and MS-5. In fact, the PA6 molecule is closely related to the hemopoiesis-supporting capacity of the stromal cells because the proliferation of HSCs was suppressed to a great extent by the anti-PA6 mAb. Affinity chromatography and mass peptide fingerprinting revealed that the protein reacting with the anti-PA6 mAb is neural cell adhesion molecule (NCAM). The frequencies of long-term cobblestone area-forming cells and long-term culture-initiating cells were significantly suppressed by repression of NCAM in the FMS/PA6-P cells using NCAM small interfering RNA. Our findings clearly indicate that NCAM functions on the maintenance of HSCs.
Wang
Xiaoli
X
First Department of Pathology, Kansai Medical University, 10-15 Fumizono-cho, Moriguchi City, Osaka 570-8506, Japan. ikehara@takii.kmu.ac.jp.
Hisha
Hiroko
H
Taketani
Shigeru
S
Inaba
Muneo
M
Li
Qiang
Q
Cui
Wenhao
W
Song
Changye
C
Fan
Tianxue
T
Cui
Yunze
Y
Guo
Kequan
K
Yang
Guoxiang
G
Fan
Hongxue
H
Lian
Zhexiong
Z
Gershwin
M Eric
ME
Ikehara
Susumu
S
D000818:Animals; D000911:Antibodies, Monoclonal / Q000494:pharmacology; D002450:Cell Communication / Q000502:physiology*; D048708:Cell Growth Processes / Q000502:physiology; D002460:Cell Line; D016207:Cytokines / Q000096:biosynthesis; D005260:Female; D006410:Hematopoiesis / Q000235:genetics / Q000502:physiology*; D051379:Mice; D008809:Mice, Inbred C3H; D008810:Mice, Inbred C57BL; D019006:Neural Cell Adhesion Molecules / Q000037:antagonists & inhibitors / Q000096:biosynthesis / Q000235:genetics / Q000502:physiology*; D011247:Pregnancy; D034741:RNA, Small Interfering / Q000235:genetics; D017154:Stromal Cells / Q000166:cytology / Q000378:metabolism / Q000502:physiology*; D014162:Transfection
D000911:Antibodies, Monoclonal; D016207:Cytokines; D019006:Neural Cell Adhesion Molecules; D034741:RNA, Small Interfering