High expression of leptin by human bone marrow adipocytes in primary culture

Adipocytes participate in the microenvironment of the bone marrow (BM), but their exact role remains to be determined. It has recently been shown that leptin, a hormone secreted from extramedullary adipocytes, could be involved in hematopoiesis. Therefore we have developed a primary culture system of human BM adipocytes to characterize their differentiation and determine whether leptin is also secreted from these adipocytes. BM cells were cultured with fetal calf and horse sera. In the presence of dexamethasone, cells with vesicles containing lipids appeared within 15 days. They expressed glycerol phosphate dehydrogenase activity and a lipolytic activity in response to isoproterenol, but expressed neither the adrenergic β3 receptor nor the mitochondrial uncoupling protein UCP1. The addition of insulin alone to the culture media did not promote adipocyte differentiation. Leptin was expressed and secreted at high levels during adipocyte differentiation. Acute exposure of differentiated adipocytes to insulin had little effect on leptin expression whereas forskolin strongly inhibited it. These results show that although human BM adipocytes differ from extramedullary adipose tissues in their sensitivity to different effectors, they are a secondary source of leptin production. They suggest that BM adipocytes could contribute to hematopoiesis via the secretion of leptin in the vicinity of hematopoietic stem cells.—Laharrague, P., Larrouy, D., Fontanilles, A‐M., Truel, N., Campfield, A., Tenenbaum, R., Galitzky, J., Corberand, J. X., Pénicaud, L., Casteilla, L. High expression of leptin by human bone marrow adipocytes in primary culture. FASEB J. 12, 733–746 (1998)

[1]  J. Gimble The function of adipocytes in the bone marrow stroma. , 1990, The New biologist.

[2]  L. Campfield,et al.  The OB Protein (Leptin) Pathway - A Link Between Adipose Tissue Mass and Central Neural Networks , 1996, Hormone and metabolic research = Hormon- und Stoffwechselforschung = Hormones et metabolisme.

[3]  J. Gimble,et al.  Adipogenesis in a murine bone marrow stromal cell line capable of supporting B lineage lymphocyte growth and proliferation: Biochemical and molecular characterization , 1990, European journal of immunology.

[4]  M. Lane,et al.  Leptin regulates proinflammatory immune responses. , 1998, FASEB journal : official publication of the Federation of American Societies for Experimental Biology.

[5]  C. Strader,et al.  Isolation of a preadipocyte cell line from rat bone marrow and differentiation to adipocytes. , 1995, Endocrinology.

[6]  H. Green,et al.  Participation of one isozyme of cytosolic glycerophosphate dehydrogenase in the adipose conversion of 3T3 cells. , 1979, The Journal of biological chemistry.

[7]  H. Sheehan,et al.  An improved method of staining leucocyte granules with Sudan black B. , 1947, The Journal of pathology and bacteriology.

[8]  L S KAPLOW,et al.  Cytochemistry of leukocyte alkaline phosphatase. Use of complex naphthol as phosphates in azo dyecoupling technics. , 1963, American journal of clinical pathology.

[9]  M. Lanotte,et al.  Histochemical and ultrastructural characteristics of a cell line from human bone-marrow stroma. , 1981, Journal of cell science.

[10]  G. Ailhaud,et al.  A preadipocyte clonal line from mouse brown adipose tissue. Short- and long-term responses to insulin and beta-adrenergics. , 1987, Experimental cell research.

[11]  M. Maffei,et al.  Positional cloning of the mouse obese gene and its human homologue , 1994, Nature.

[12]  M. Berlan,et al.  In situ assessment of the role of the β1, β2‐ and β3‐adrenoceptors in the control of lipolysis and nutritive blood flow in human subcutaneous adipose tissue , 1996 .

[13]  J. Friedman,et al.  Expression of ob Gene in Adipose Cells , 1996, The Journal of Biological Chemistry.

[14]  A. Kosaki,et al.  Reduced Expression of the Leptin Gene (ob) by Catecholamine Through a GS Protein-Coupled Pathway in 3T3-L1 Adipocytes , 1996, Diabetes.

[15]  M. Maffei,et al.  Positional cloning of the mouse obese gene and its human homologue , 1995, Nature.

[16]  N. Petrakis Some physiological and developmental considerations of the temperature-gradient hypothesis of bone marrow distribution. , 1966, American journal of physical anthropology.

[17]  J. Gimble,et al.  Osteoblastic gene expression during adipogenesis in hematopoietic supporting murine bone marrow stromal cells , 1993, Journal of cellular physiology.

[18]  M. Chiesi,et al.  Regulation of ob gene mRNA levels in cultured adipocytes , 1996, FEBS letters.

[19]  A. Strosberg,et al.  The human β3-adrenoceptor: the search for a physiological function , 1994 .

[20]  J. Triffitt,et al.  Adipocytic cells cultured from marrow have osteogenic potential. , 1991, Journal of cell science.

[21]  L. Kaplow SIMPLIFIED MYELOPEROXIDASE STAIN USING BENZIDINE DIHYDROCHLORIDE. , 1965, Blood.

[22]  C. Li,et al.  ESTERASES IN HUMAN LEUKOCYTES , 1973, The journal of histochemistry and cytochemistry : official journal of the Histochemistry Society.

[23]  Su‐Li Cheng,et al.  Expression of bone matrix proteins during dexamethasone‐induced mineralization of human bone marrow stromal cells , 1996, Journal of cellular biochemistry.

[24]  K. Nakao,et al.  Human Obese Gene Expression: Adipocyte-Specific Expression and Regional Differences in the Adipose Tissue , 1995, Diabetes.

[25]  H. Snodgrass,et al.  Leptin stimulates fetal and adult erythroid and myeloid development. , 1997, Blood.

[26]  L. Weiss,et al.  Seasonal variations in hematopoiesis in the dermal bones of the nine‐banded armadillo , 1956, The Anatomical record.

[27]  C. Li,et al.  STUDIES OF ACID PHOSPHATASE ISOENZYMES IN HUMAN LEUKOCYTES , 1970, The journal of histochemistry and cytochemistry : official journal of the Histochemistry Society.

[28]  A. Bathija,et al.  Marrow adipose tissue: Response to erythropoiesis , 1978, American journal of hematology.

[29]  M. Wabitsch,et al.  Promoting effect of glucocorticoids on the differentiation of human adipocyte precursor cells cultured in a chemically defined medium. , 1989, The Journal of clinical investigation.

[30]  L. Lajtha,et al.  Conditions controlling the proliferation of haemopoietic stem cells in vitro , 1977, Journal of cellular physiology.

[31]  L. Casteilla,et al.  Evidence for numerous brown adipocytes lacking functional beta 3-adrenoceptors in fat pads from nonhuman primates. , 1996, The Journal of clinical endocrinology and metabolism.

[32]  D. Mason,et al.  Alkaline phosphatase and peroxidase for double immunoenzymatic labelling of cellular constituents. , 1978, Journal of clinical pathology.

[33]  L Berga,et al.  Age-related variations of fat tissue fraction in normal human bone marrow depend both on size and number of adipocytes: a stereological study. , 1989, Experimental hematology.

[34]  J. Gimble,et al.  The function of adipocytes in the bone marrow stroma: an update. , 1996, Bone.

[35]  R. Oreffo,et al.  Modulation of osteogenesis and adipogenesis by human serum in human bone marrow cultures. , 1997, European journal of cell biology.

[36]  L. Kaplow,et al.  Brief Report: Simplified Myeloperoxidase Stain Using Benzidine Dihydrochloride , 1965 .

[37]  W. Alexander,et al.  Leptin can induce proliferation, differentiation, and functional activation of hemopoietic cells. , 1996, Proceedings of the National Academy of Sciences of the United States of America.

[38]  M. Mattei,et al.  Human uncoupling protein gene: Structure, comparison with rat gene, and assignment to the long arm of chromosome 4 , 1990, Journal of cellular biochemistry.

[39]  P. Chomczyński,et al.  Single-step method of RNA isolation by acid guanidinium thiocyanate-phenol-chloroform extraction. , 1987, Analytical biochemistry.

[40]  J. Falkenburg,et al.  Regulation of myelopoiesis by murine fibroblastic and adipogenic cell lines. , 1993, Experimental hematology.

[41]  B. Bennett,et al.  A role for leptin and its cognate receptor in hematopoiesis , 1996, Current Biology.