Primary Mouse Bone Marrow-derived Cells Research Articles

Antipostmenopausal effects of Stauntonia hexaphylla and Vaccinium bracteatum fruit combination in estrogen-deficient rats.

BackgroundClimacterium is a series of physical and mental symptoms occurring in women and men due to decreased levels of sex hormones. Women lose the ability to become pregnant due to decreased ovarian estrogen production; the initial symptom being hot flushes. In addition, urogenital atrophy, sexual dysfunction, mood changes, and osteoporosis occur. Extracts of Stauntonia hexaphylla (SH) and Vaccinium bracteatum (VB) fruits, with a wide range of biological activities, are widely used in traditional herbal medicine.ObjectiveThe purpose of this study was to investigate the mitigation of menopausal symptoms, such as hot flushes and postmenopausal osteoporosis after combinatorial treatment with SH and VB (SHVB) of ovariectomized (OVX) rats.DesignWe measured the bone regenerative effect of SHVB on receptor activator of nuclear factor-κB (NF-κB) ligand-induced osteoclast differentiation in vitro and on ovariectomy-induced osteoporosis in vivo. We investigated the effect of SHVB in a rat model of menopausal hot flushes, in which the tail skin temperature increases following ovariectomy-induced rapid decline in estrogen levels.ResultsSHVB inhibited osteoclast formation and tartrate-resistant acid phosphatase activity in primary mouse bone marrow-derived cells. In an estrogen deficiency-induced rat model, measurement of serum bone turnover factors showed that treatment with SHVB lowered the increased bone turnover. Additionally, SHVB decreased OVX-induced bone loss of the total femur. SHVB inhibited osteoclast differentiation, prevented bone mass reduction, and improved trabecular bone structure and biochemical markers in OVX-induced osteoporosis. In addition, administration of SHVB significantly ameliorated the changes in skin temperature in OVX rats.ConclusionSHVB improved the symptoms of menopause. These results provide the foundation for developing SHVB as a natural substance to replace hormones in the future.

Transfection of bone marrow derived cells with immunoregulatory proteins

In vitro electroporation gene transfer was first performed in 1982. Today, this technology has become one of the major vehicles for non-viral transfection of cells. All non-viral transfections, such as calcium phosphate precipitation, lipofection, and magnetic transfection, have been shown to achieve a transfection efficiency of up to 70% in commonly used cell lines, but not in primary cells. Here we describe the use of electroporation to transfect primary mouse bone marrow-derived cells, such as macrophages (Mφ) and dendritic cells (DCs) with high efficiencies (45%–72%) and minimal cell death. The transfection efficiencies and cell death varied depending on the culture duration of the DCs and Mφ. Moreover, the electroporation efficiency was increased when conditioning medium was used for culturing the cells. Furthermore, we demonstrated that measuring the plasmid-encoded secreted proteins is a highly sensitive method for determining the transfection efficiency. In summary, electroporation with plasmid vectors is an efficient method for producing DCs and Mφ with transient expression of immunoregulatory proteins.

Bone regenerative effect of aqueous Cynanchum wilfordii extract in receptor activator of nuclear factor-κB ligand-induced osteoclast differentiation and estrogen deficiency-induced osteoporosis

<p>Osteoporosis increases with age, most frequently in postmenopausal women because of reduced ovarian hormone levels. Furthermore, estrogen deficiency impairs trabecular metaphyseal bone. Although efficacious, long-term hormone replacement therapy (HRT) has estrogen-like side effects including breast and endometrial cancers, and non-hormonal or herbal therapies may be safer alternatives. Therefore, the aim of this study was to investigate the effects of aqueous extracts of <em>Cynanchum wilfordii</em> (CWW) on receptor activator of nuclear factor-κ B (NF-κ B) ligand (RANKL)-induced osteoclast differentiation <em>in vitro</em> and ovariectomy-mediated osteoporosis <em>in vivo</em>. CWW inhibited RANKL-induced osteoclast formation and tartrate-resistant acid phosphatase (TRAP) activity in primary mouse bone marrow-derived cells. We investigated the osteoprotective effect of CWW in an ovariectomized (OVX) Sprague-Dawley rat model treated with vehicle (OVX/vehicle), 17<span style="font-family: 맑은 고딕;">β</span>-estradiol (OVX/E2), or three CWW doses (100, 200, and 400 mg/kg). After a 24-week treatment, the body and uterus weights were not affected except in the OVX/E2 group. Additionally, bone mineral density (BMD) and histological analyses showed that the BMD of the femurs of CWW400-treated rats was significantly higher than that of the OVX/vehicle rats, and comparable to that of the OVX/E2 group rats. Serum levels of bone turnover markers alkaline phosphatase (ALP), osteocalcin, collagen type I C-telopeptide, and TRAP significantly decreased in the CWW400 group. Our results show that compared to the vehicle, CWW had a significant anti-osteoporotic effect in the OVX model. Taken together, CWW exhibited inhibitory effects on osteoclastogenesis <em>in vitro,</em> and we confirmed its in vivo efficacy in the prevention of osteoporosis.</p>