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Constitutive and modulated cytokine expression in two permanent human bone marrow stromal cell lines.
Exp Hematol. 1996 Jan; 24(1):1-10.EH

Abstract

We present a detailed analysis of cytokine expression patterns of the two permanent human bone marrow stromal cell lines, L87/4 and L88/5. These cell lines, previously established in our laboratory, are highly radiotolerant without cell detachment and support long-term cultures of CD(34+)-enriched human cord blood cells. RT-PCR analysis of 22 different cytokines or cytokine receptor mRNAs showed an almost identical expression pattern in the two stromal cell lines compared to primary human Dexter-type stroma. Since stromal feeder lines employed in long-term cultures usually are irradiated and grown in media containing corticosteroids, we analyzed the impact of irradiation and dexamethasone on cytokine production in the two cell lines by RT-PCR, Northern blot analysis, bioassays, and RIAs. By RT-PCR analysis, constitutive mRNA expression of c-kit, G-CSF, GM-CSF, IL-1 beta, IL-6, IL-7, IL-8, IL-11, Kit ligand (KL), LIF, M-CSF, MIP-1 alpha, TGF-beta, and TNF-alpha was demonstrated in both cell lines, with L87/4 a more potent cytokine producer than L88/5. Northern blot data showed an increase in mRNA levels for GM-CSF, IL-1 beta, and LIF by irradiation and IL-1 alpha treatment in both cell lines. IL-1 alpha-induced GM-CSF, IL-1 beta, IL-6, IL-11, and LIF mRNA levels were reduced by the addition of dexamethasone, whereas dexamethasone had no influence on the amounts of IL-1 alpha-induced G-CSF mRNA. L87/4 and, to a lower extent, L88/5 cells showed dexamethasone-dependent increases in KL mRNA, while KL mRNA levels were not stimulated by IL-1 alpha.

Authors+Show Affiliations

Thalmeier K
GSF-Institut für Experimentelle Hämatologie, München, Germany.
Meissner P
No affiliation info available
Reisbach G
No affiliation info available
Hültner L
No affiliation info available
Mortensen BT
No affiliation info available
Brechtel A
No affiliation info available
Oostendorp RA
No affiliation info available
Dörmer P
No affiliation info available

MeSH

Base SequenceBlotting, NorthernBone MarrowBone Marrow CellsCell LineCytokinesDexamethasoneGene ExpressionGranulocyte-Macrophage Colony-Stimulating FactorGrowth InhibitorsHumansInterleukin-1Interleukin-11Interleukin-6Leukemia Inhibitory FactorLymphokinesMolecular Sequence DataPolymerase Chain ReactionRNA, MessengerRNA-Directed DNA PolymeraseStromal Cells

Pub Type(s)

Comparative Study
Journal Article

Language

eng

PubMed ID

8536785

Citation

Thalmeier, K, et al. "Constitutive and Modulated Cytokine Expression in Two Permanent Human Bone Marrow Stromal Cell Lines." Experimental Hematology, vol. 24, no. 1, 1996, pp. 1-10.
Thalmeier K, Meissner P, Reisbach G, et al. Constitutive and modulated cytokine expression in two permanent human bone marrow stromal cell lines. Exp Hematol. 1996;24(1):1-10.
Thalmeier, K., Meissner, P., Reisbach, G., Hültner, L., Mortensen, B. T., Brechtel, A., Oostendorp, R. A., & Dörmer, P. (1996). Constitutive and modulated cytokine expression in two permanent human bone marrow stromal cell lines. Experimental Hematology, 24(1), 1-10.
Thalmeier K, et al. Constitutive and Modulated Cytokine Expression in Two Permanent Human Bone Marrow Stromal Cell Lines. Exp Hematol. 1996;24(1):1-10. PubMed PMID: 8536785.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Constitutive and modulated cytokine expression in two permanent human bone marrow stromal cell lines. AU - Thalmeier,K, AU - Meissner,P, AU - Reisbach,G, AU - Hültner,L, AU - Mortensen,B T, AU - Brechtel,A, AU - Oostendorp,R A, AU - Dörmer,P, PY - 1996/1/1/pubmed PY - 1996/1/1/medline PY - 1996/1/1/entrez SP - 1 EP - 10 JF - Experimental hematology JO - Exp Hematol VL - 24 IS - 1 N2 - We present a detailed analysis of cytokine expression patterns of the two permanent human bone marrow stromal cell lines, L87/4 and L88/5. These cell lines, previously established in our laboratory, are highly radiotolerant without cell detachment and support long-term cultures of CD(34+)-enriched human cord blood cells. RT-PCR analysis of 22 different cytokines or cytokine receptor mRNAs showed an almost identical expression pattern in the two stromal cell lines compared to primary human Dexter-type stroma. Since stromal feeder lines employed in long-term cultures usually are irradiated and grown in media containing corticosteroids, we analyzed the impact of irradiation and dexamethasone on cytokine production in the two cell lines by RT-PCR, Northern blot analysis, bioassays, and RIAs. By RT-PCR analysis, constitutive mRNA expression of c-kit, G-CSF, GM-CSF, IL-1 beta, IL-6, IL-7, IL-8, IL-11, Kit ligand (KL), LIF, M-CSF, MIP-1 alpha, TGF-beta, and TNF-alpha was demonstrated in both cell lines, with L87/4 a more potent cytokine producer than L88/5. Northern blot data showed an increase in mRNA levels for GM-CSF, IL-1 beta, and LIF by irradiation and IL-1 alpha treatment in both cell lines. IL-1 alpha-induced GM-CSF, IL-1 beta, IL-6, IL-11, and LIF mRNA levels were reduced by the addition of dexamethasone, whereas dexamethasone had no influence on the amounts of IL-1 alpha-induced G-CSF mRNA. L87/4 and, to a lower extent, L88/5 cells showed dexamethasone-dependent increases in KL mRNA, while KL mRNA levels were not stimulated by IL-1 alpha. SN - 0301-472X UR - https://www.unboundmedicine.com/medline/citation/8536785/Constitutive_and_modulated_cytokine_expression_in_two_permanent_human_bone_marrow_stromal_cell_lines_ L2 - https://web.expasy.org/cellosaurus/CVCL_6837 DB - PRIME DP - Unbound Medicine ER -
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