Isolation and hormonal responsiveness of primary cultures of human bone-derived cells: gender and age differences

Katzburg, Sara; Lieberherr, Michèle; Ornoy, Asher; Klein, Benjamin Y.; Hendel, David; Sömjen, Dalia

doi:10.1016/s8756-3282(99)00225-2

Cited by 87 publications

(85 citation statements)

References 41 publications

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“…Other reports demonstrated that osteogenic progenitor cells were maintained during aging [47][48][49]. The question of changes in the osteoprogenitor cell population with age is further complicated by the possibility of sex-dependent differences [50][51][52]. Age-related osteoponia may also depend from the skeletal site, depending to the variable trabecular-cortical bone ratio at different locations [53].…”

Section: Discussionmentioning

confidence: 99%

Fetal bone cells for tissue engineering

Montjovent

Burri

Mark

et al. 2004

Bone

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We envision the use of human fetal bone cells for engineered regeneration of adult skeletal tissue. A description of their cellular function is then necessary. To our knowledge, there is no description of human primary fetal bone cells treated with differentiation factors. The characterization of fetal bone cells is particularly important as the pattern of secreted proteins from osteoblasts has been shown to change during aging. In the first part of this work, human primary fetal bone cells were compared to adult bone cells and mesenchymal stem cells for their ability to proliferate and to differentiate into osteoblasts in vitro. Cell proliferation, gene expression of bone markers, alkaline phosphatase (ALP) activity, and mineralization were analyzed during a time-course study. In the second part of this paper, bone fetal cells behavior exposed to osteogenic factors is further detailed. The doubling time of fetal bone cells was comparable to mesenchymal stem cells but significantly shorter than for adult bone cells. Gene expression of cbfa-1, ALP, a1 chain of type I collagen, and osteocalcin were upregulated in fetal bone cells after 12 days of treatment, with higher inductions than for adult and mesenchymal stem cells. The increase of ALP enzymatic activity was stronger for fetal than for adult bone cells reaching a maximum at day 10, but lower than for mesenchymal stem cells. Importantly, the mineralization process of bone fetal cells started earlier than adult bone and mesenchymal stem cells. Proliferation of fetal and adult bone cells was increased by dexamethasone, whereas 1a,25-dihydroxyvitamin D 3 did not show any proliferative effect. Mineralization studies clearly demonstrated the presence of calcium deposits in the extracellular matrix of fetal bone cells. Nodule formation and calcification were strongly increased by the differentiation treatment, especially by dexamethasone. This study shows for the first time that human primary fetal bone cells could be of great interest for bone research, due to their fast growth rate and their ability to differentiate into mature osteoblasts. They represent an interesting and promising potential for therapeutic use in bone tissue engineering.

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Section: Discussionmentioning

confidence: 99%

Fetal bone cells for tissue engineering

Montjovent

Burri

Mark

et al. 2004

Bone

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“…1B). Therefore, these easily obtained and well characterized osteoblast cultures can be made estrogen responsive in vitro, replicating the hormone-sensitive status of osteoblasts that occur in more mature organisms in vivo (43)(44)(45)(46). Importantly, this osteoblast cell culture model provides a sensitive system to examine specific aspects of ER␣ on osteoblast activity uncomplicated by the presence of endogenous ERs.…”

Section: Er-dependent Gene Expression In Culturedmentioning

confidence: 99%

“…We chose to avoid possible complications from antibiotic toxicity, unknown effects from stable gene integration, and phenotypic drift by cells in continuous culture with our approach. Even so, it is difficult to know how the level of ER␣ expression in any transfected cell model compares with that in adult bone because, even in vivo, levels of ER␣ vary considerably with age and with anatomical bone location (43)(44)(45)(46). However, expression of ER␣ by transient gene transfection seemed appropriate and predictable, because the ability of estrogen to driven reporter gene expression by ERE was steroid- FIG.…”

Section: Fig 3 Er Associates With Runx2mentioning

confidence: 99%

Runx2 Integrates Estrogen Activity in Osteoblasts

McCarthy

Chang

Liu

et al. 2003

Journal of Biological Chemistry

106

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Steroids significantly effect skeletal integrity. For example, bone mass decreases with glucocorticoid excess or with estrogen depletion after menopause. Glucocorticoid suppresses gene expression by an essential skeletal tissue transcription factor, Runx2, in rat osteoblasts. We now report that estrogen enhances Runx2 activity in dose-and estrogen receptor-dependent ways independently of changes in Runx2 levels or its DNA binding potential. Estrogen receptor and Runx2 can be collected by co-immunoprecipitation. By two-hybrid gene expression analysis, high affinity complex formation involves portions of Runx2 outside of its own DNA binding domain and the DNA binding domain of the estrogen receptor. Consistent with this interaction, the stimulatory effect of estrogen on Runx2 activity is lost when the DNA binding domain of the estrogen receptor is eliminated. Unlike the stimulatory effect of estrogen and the inhibitory effect of glucocorticoid, androgen fails to increase Runx2 activity, whereas Runx2 potently suppresses gene expression induced by all three steroids. Finally, estrogen increases gene transcription by the transforming growth factor-␤ type I receptor gene promoter, which contains several Runx binding sequences, and enhances Smad dependent gene expression by transforming growth factor-␤ in osteoblasts. These results reveal that Runx2 can integrate complex effects on gene transcription in hormone-, growth factor-, and tissue-restricted ways.

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“…Diabetes has been associated with a net loss of bone [17,18], with reduction of new bone formation and decreased bone mineral density. In diabetic mice the up-regulation of specific transcription factors is attenuated, resulting in deficiency in conversion of mesenchymal cells to osteoblasts [17,18].…”

Section: Introductionmentioning

confidence: 99%

“…In diabetic mice the up-regulation of specific transcription factors is attenuated, resulting in deficiency in conversion of mesenchymal cells to osteoblasts [17,18].…”

Section: Introductionmentioning

confidence: 99%

Phytoestrogenic Compounds and Their Synthetic Analogs, Contrary to Estradiol- 17b Stimulates Human Derived Female Cultured Bone Cells in Hyperglycemic Conditions

Sömjen¹,

Katzburg²,

Tamir³

et al. 2011

J Steroids Hormon Sci

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Cultured female-derived human osteoblasts (hObs) responded by different parameters to the phytoestrogens: daidzein (D), glabrene (Gla) and glabridin (Glb), to their synthetic derivatives; carboxy-daidzein (cD) and to estradiol-17β (E 2 ). Since the skeletal protective effects of estrogens are not discernible in diabetic women, we tested the effects of these compounds on hObs grown in growth medium with high glucose (HG; 9.0g/L; 44mM) compared to normal glucose (NG; 4.5g/L; 22mM) using the stimulation of creatine kinase specific activity (CK) and 3 [H] thymidine incorporation into DNA (DNA) as hormonal responsiveness markers. HG slightly increased DNA and CK in hObs.Stimulations by E 2 was abolished and by cD and D was slightly decreased in HG, but not by Gla and Glb in both age groups. Growing hObs in HG upregulated the expression of mRNA of both ER and ERβ in cells from pre-but not from post-menopausal women. Cells from both age groups express also mRNA for 25 hydroxy vitamin D 3 1-α hydroxylase and showed enzymic activity which were down-regulated by HG in both age groups. Whether Gla and Glb act differentially via ERs and/or 1-α hydroxylase is not yet established.Since these compounds are active even in HG, they might be used for treating hyperglycemic/diabetic women. Journal of Steroids & Hormonal Science Materials and Methods ReagentsAll reagents used were analytical grade. Estradiol-17β (E 2 ), daidzein (D) and the creatine kinase (CK) assay kit were purchased from Sigma Chemicals Co. (St. Louis, MO). Carboxy-D (cD) was synthesized by us [20], licorice products: Gla and Glb were produced by us [19]. Cell culturesHuman bones were obtained from biopsies of patients undergoing corrective surgery following accidental injury, hip or knee replacement. All patients (women and men) were healthy, non-osteoporotic and not receiving hormonal replacement treatment. Three groups were defined: Pre-menopausal women, ranging between 37-55 years old, (n=5). Post-menopausal women, ranging between 60-84 years old, (n=5). The non-enzymic method for isolation and culture of human bone cells and their characterization as osteoblasts was described previously [12]. Briefly, samples of the trabecular surface of the iliac crest or long bones were cut into 1mm 3 pieces and extensively and repeatedly washed with phosphate buffered saline (PBS) to remove blood components. The explants, with no enzymatic digestion, were seeded in 100mm diameter tissue culture dishes and incubated in DMEM medium without Ca ++ (to avoid fibroblastic growth [12], containing 10% fetal calf serum (FCS) and antibiotics. Cell outgrowth from the bone explants was apparent after 6-10 days. First passage cells were seeded at a density of 3x10 5 cells per 35mm tissue culture dish in phenol red free DMEM with 10% charcoal stripped FCS and incubated at 37°C in 5% CO 2 . To obtain "high glucose" (HG) conditions, the medium including the FCS, was supplemented with glucose up to a final concentration of 44nM (9.0gm/ liter). Glucose concentration in the regula...

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Isolation and hormonal responsiveness of primary cultures of human bone-derived cells: gender and age differences

Cited by 87 publications

References 41 publications

Fetal bone cells for tissue engineering

Fetal bone cells for tissue engineering

Runx2 Integrates Estrogen Activity in Osteoblasts

Phytoestrogenic Compounds and Their Synthetic Analogs, Contrary to Estradiol- 17b Stimulates Human Derived Female Cultured Bone Cells in Hyperglycemic Conditions

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