Bone marrow stromal proteoglycan heterogeneity: Phenotypic variability between cell lines and the effects of glucocorticoid
✍ Scribed by Stuart A. Bentley; Suzanne L. Kirby; Pervin Anklesaria; Joel S. Greenberger
- Publisher
- John Wiley and Sons
- Year
- 1988
- Tongue
- English
- Weight
- 656 KB
- Volume
- 136
- Category
- Article
- ISSN
- 0021-9541
No coin nor oath required. For personal study only.
✦ Synopsis
Hematopoiesis in vivo is dependent upon the interaction of hematopoietic stem cells with a complex microenvironment, of which stromal proteoglycans are an important functional component. Certain bone marrow stromal cell lines provide a microenvironment that supports hematopoiesis in vitro, a function that is dependent upon glucocorticoid supplementation. Proteoglycan synthesis in the hematopoietic-supportive D2XRI1, B16 and 14F1 bone marrow stromal cell lines was studied by '"S-sulfate precursor labelling and ion-exchange separation, followed by isopyknic CsCl density centrifugation and gel filtration HPLC. The effects of glucocorticoid were also investigated. A similar pattern of proteoglycan heterogeneity was observed in all three cell lines, although there was considerable quantitative variation. All cultures synthesized three species of chondroitini dermatan sulfate (CSIDS) proteoglycans: DS1, excluded from a Bio-Sil TSK-400 HPLC column, and DS2, eluting at Kd = 0.31, were present mainly in the culture media. The smallest (DS3) eluted at Kd = 0.63 and was present mainly in the cell layers. CSiDS species were the major proteoglycans in all cultures. Hydrocortisone-free cultures also synthesized heparan sulfate (HS) proteoglycans, including a cell-associated form (HSl), partially excluded from the TSK-400 column, and a secretory form (HS2), eluting at Kd = 0.15. DZXRII cells also secreted an apparently-unique, high-density proteoglycan, Kd = 0.65, into the culture medium. Hydrocortisone at lo-" M virtually abolished HS proteoglycan synthesis in all three cell lines, and altered the pattern of CSiDS proteoglycans in the culture media, increasing the quantity of DS1 and DS3, and reducing the quantity of DS2.