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

探讨人脐带单个核细胞（uMNC）诱导破骨细胞的最佳培养条件。在含10%胎牛血清（FBS）、20%马血清（HS）或2%牛血清白蛋白（BSA）的DMEM/F12培养基中培养uMNC，加入或不加入M-CSF、IL-4、破骨细胞分化因子和1 α，25-二羟维生素D3（VD）。在存在VD的情况下，在用添加FBS的M-GSF或添加BSA的M-CSF培养的uMNC中，或在存在IL-4的情况下，在用添加HS的M-CSF培养的uMNC中，有效地诱导了抗酒石酸盐酸性磷酸酶阳性的多核细胞。在存在VD的情况下用添加了HS的M-CSF或存在IL-4的情况下用添加了BSA的M-CSF培养的uMNC中，降钙素诱导的cAMP产生显著增加。在存在VD的情况下，用添加了HS的M-CSF培养的uMNC中的骨吸收活性增加。E_2不能刺激含有M-CSF和IL-4的BSA培养的uMNC产生cAMP。研究了在有（+）或无（-）E2的情况下，用含有M-CSF和IL-4的BSA培养的具有雌激素受体基因多态性PP、Pp和pp的3 uMNC中LPS诱导的TNFα产生。LPS诱导的E2（+）uMNC-/- E2（-）uMNC产生TNFα的比值，PP为0.60±0.06，Pp为1.06±0.06，pp为1.44±0.12。因此，雌激素及其受体基因多态性均影响LPS诱导的UMNC TNFα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.