Direct Effect of Estrogen on Cultured Osteoclast Derived from Umbilical Cord Blood Mononuclear Cell

探讨了人脐血单个核细胞(UMNC)诱导破骨细胞的最佳培养条件。在含10%胎牛血清、20%马血清和2%牛血清白蛋白的DMEM/F12培养液中，分别加入或不加入下列物质：M-CSF、IL-4、破骨细胞分化因子和1-α，25-二羟基维生素D3(VD)。抗酒石酸酸性磷酸酶阳性的uMNC在M-GSF+FBS或M-CSF+BSA+VD或M-CSF+HS+IL-4培养的uMNC中均能被有效地诱导。在血管性痴呆或IL-4存在的情况下，M-CSF加HS或M-CSF加牛血清白蛋白培养的uMNC均可明显增加降钙素诱导的cAMP生成。在血管性痴呆存在的情况下，加入M-CSF和HS培养的uMNC的骨吸收活性增加。雌激素(E_2)不能刺激含有M-CSF和IL-4的牛血清白蛋白培养的uMNC产生cAMP。用含有M-α和IL-4的牛血清白蛋白培养3uMNC，观察雌激素受体基因多态性PP、PP和PP在E2(+)和不加E2(-)条件下诱导的肿瘤坏死因子mRNA的产生。脂多糖诱导的E_2(+)uMNC-与E_2(-)uMNC产生肿瘤坏死因子α的比值分别为0.60±0.06、1.06±0.06和1.44±0.12。因此，雌激素及其受体基因多态性均影响内毒素诱导的巨噬细胞肿瘤坏死因子α的产生。

Optimal culture condition was assessed in inducing osteoclast from the human umbilical cord mononuclear cells (uMNC). The uMNC were cultured in three DMEM/F12 medium containing 10% fetal bovine serum (FBS), 20% horse serum (HS) or 2% BSA(BSA), with or without following substances ; M-CSF, IL-4, osteoclast differentiation factor, and 1 α, 25-dihydroxyvitamine D3 (VD). Multinucleated cells positive for tartrate-resistant acid phosphatase were efficiently induced in uMNC cultured with M-GSF added FBS or M-CSF added BSA in the presence of VD, or with M-CSF added HS in the presence of IL-4. Calcitonin-induced cAMP production was significantly increased in uMNC cultured with M-CSF added HS in the presence of VD or M-CSF added BSA in the presence of IL-4. Bone resorbing activity was increased in uMNC cultured with M-CSF added HS in the presence of VD. Estradiol (E2) did not stimulate cAMP production of in uMNC cultured with BSA containing M-CSF and IL-4. LPS-induced TNFα production was investigated in 3 uMNC with estrogen receptor gene polymorphism PP, Pp, and pp cultured with BSA containing M-CSF and IL-4 in the precence (+) or absence of E2 (-). The LPS-induced TNFα production ratio of E2 (+) uMNC- to - E2 (-) uMNC was 0.60±0.06 for uMNC with PP, 1.06±0.06 for uMNCwith Pp, and 1.44±0.12 for uMNC with pp. Thus, both the estrogen and its receptor gene polymorphism affected the LPS-induced TNFα production in UMNC.