Distinct hematopoietic support by two human stromal cell lines.
OBJECTIVE
The hematopoietic microenvironment is complex, and the role of myofibroblast in its function is crucial. In order to obtain a stable model reflecting this particular cell type, we have previously established human bone marrow cell lines from primary myofibroblastic Stro1(+) population (pStro1(+)). We placed HPV16 E6 and E7 expression under the control of different promoters. Here, we have characterized and studied the hematopoietic support for two cell lines corresponding to the promoters alpha-SM (alphaSM-56 line) and SV40 (SV40-56 line).


METHODS
The expression profile was analyzed at the RNA level by gene array and at the protein level by Western blot, flow cytometry, and ELISA. Hematopoietic support determined using colony-forming unit (CFU) and stroma-adherent colony-forming cell (SA-CFC) assays.


RESULTS
The phenotype of cell lines was not significantly modified compared with primary myofibroblastic cells. They secreted a broad spectrum of hematopoietic cytokines and nonspecific mediators. The two lines allowed the growth of hematopoietic precursors and had different support capabilities.


CONCLUSIONS
We have extensively characterized two novel human bone marrow stromal cell lines. They retained a myofibroblastic phenotype and have substantial but different hematopoietic support capabilities. These lines provided a basis for determining stromal factors involved in stem-cell regulation.
Loeuillet
C
C
EFS de Bourgogne-Franche Comté, Besançon, France. c.loeuillet@voila.fr
Bernard
G
G
Rémy-Martin
J
J
Saas
P
P
Hervé
P
P
Douay
L
L
Chalmers
D
D
D018952:Antigens, CD34 / Q000032:analysis; D001854:Bone Marrow Cells / Q000166:cytology; D002448:Cell Adhesion; D015815:Cell Adhesion Molecules / Q000032:analysis; D002460:Cell Line; D002478:Cells, Cultured; D018920:Coculture Techniques; D003114:Colony-Forming Units Assay; D015925:Cryopreservation; D003470:Culture Media; D016207:Cytokines / Q000032:analysis / Q000235:genetics*; D005434:Flow Cytometry; D006412:Hematopoietic Stem Cells / Q000166:cytology / Q000502:physiology*; D006801:Humans; D016023:Integrins / Q000032:analysis; D010641:Phenotype; D017154:Stromal Cells / Q000166:cytology / Q000502:physiology*
D018952:Antigens, CD34; D015815:Cell Adhesion Molecules; D003470:Culture Media; D016207:Cytokines; D016023:Integrins