中华血液学杂志
中華血液學雜誌
중화혈액학잡지
Chinese Journal of Hematology
2014年
7期
623-627
,共5页
赵思捷%付蓉%刘惠%王一浩%李丽娟%刘春燕%张田%丁少雪%高珊
趙思捷%付蓉%劉惠%王一浩%李麗娟%劉春燕%張田%丁少雪%高珊
조사첩%부용%류혜%왕일호%리려연%류춘연%장전%정소설%고산
CCL3%骨髓瘤骨病%成骨细胞
CCL3%骨髓瘤骨病%成骨細胞
CCL3%골수류골병%성골세포
CCL3%Myeloma bone disease%Osteoblast
目的 建立多发性骨髓瘤(MM)患者成骨细胞体外培养体系,检测成骨细胞膜CCL3受体(CCR1)表达,研究CCL3对MM患者成骨细胞(OB)的作用.方法 采用密度梯度离心法分离骨髓单个核细胞(BMMNC),应用地塞米松、β-甘油磷酸钠及维生素C诱导并纯化获得MM患者成骨细胞,以碱性磷酸酶染色及Von Kossa's钙染色鉴定成骨细胞,用流式细胞术(FCM)测定培养后成骨细胞纯度.采用FCM分别检测成骨细胞膜CCR1的表达水平,并在培养体系中加入CCL3干预后比较成骨细胞数量、形态及钙质沉积的变化.结果 经联合诱导培养获得的细胞形态不规则,有较多突起,碱性磷酸酶染色及Von Kossa's钙染色均为阳性,提示培养获得的细胞为成骨细胞,经FCM测定纯度均在90％以上.MM患者成骨细胞膜CCR1的表达水平为(74.48±7.31)％,高于正常对照组[(48.35±8.81)％](P＜0.05).加入CCL3干预后,MM患者成骨细胞形态及数量变化不显著,Von Kossa's钙染示钙结节明显减少.结论 采用骨髓来源的单个核细胞可用于体外培养MM患者成骨细胞.MM患者成骨细胞膜高表达CCR1,CCL3可抑制成骨细胞功能.提示CCL3可能在MM骨病成骨受抑机制中发挥一定作用.
目的 建立多髮性骨髓瘤(MM)患者成骨細胞體外培養體繫,檢測成骨細胞膜CCL3受體(CCR1)錶達,研究CCL3對MM患者成骨細胞(OB)的作用.方法 採用密度梯度離心法分離骨髓單箇覈細胞(BMMNC),應用地塞米鬆、β-甘油燐痠鈉及維生素C誘導併純化穫得MM患者成骨細胞,以堿性燐痠酶染色及Von Kossa's鈣染色鑒定成骨細胞,用流式細胞術(FCM)測定培養後成骨細胞純度.採用FCM分彆檢測成骨細胞膜CCR1的錶達水平,併在培養體繫中加入CCL3榦預後比較成骨細胞數量、形態及鈣質沉積的變化.結果 經聯閤誘導培養穫得的細胞形態不規則,有較多突起,堿性燐痠酶染色及Von Kossa's鈣染色均為暘性,提示培養穫得的細胞為成骨細胞,經FCM測定純度均在90％以上.MM患者成骨細胞膜CCR1的錶達水平為(74.48±7.31)％,高于正常對照組[(48.35±8.81)％](P＜0.05).加入CCL3榦預後,MM患者成骨細胞形態及數量變化不顯著,Von Kossa's鈣染示鈣結節明顯減少.結論 採用骨髓來源的單箇覈細胞可用于體外培養MM患者成骨細胞.MM患者成骨細胞膜高錶達CCR1,CCL3可抑製成骨細胞功能.提示CCL3可能在MM骨病成骨受抑機製中髮揮一定作用.
목적 건립다발성골수류(MM)환자성골세포체외배양체계,검측성골세포막CCL3수체(CCR1)표체,연구CCL3대MM환자성골세포(OB)적작용.방법 채용밀도제도리심법분리골수단개핵세포(BMMNC),응용지새미송、β-감유린산납급유생소C유도병순화획득MM환자성골세포,이감성린산매염색급Von Kossa's개염색감정성골세포,용류식세포술(FCM)측정배양후성골세포순도.채용FCM분별검측성골세포막CCR1적표체수평,병재배양체계중가입CCL3간예후비교성골세포수량、형태급개질침적적변화.결과 경연합유도배양획득적세포형태불규칙,유교다돌기,감성린산매염색급Von Kossa's개염색균위양성,제시배양획득적세포위성골세포,경FCM측정순도균재90％이상.MM환자성골세포막CCR1적표체수평위(74.48±7.31)％,고우정상대조조[(48.35±8.81)％](P＜0.05).가입CCL3간예후,MM환자성골세포형태급수량변화불현저,Von Kossa's개염시개결절명현감소.결론 채용골수래원적단개핵세포가용우체외배양MM환자성골세포.MM환자성골세포막고표체CCR1,CCL3가억제성골세포공능.제시CCL3가능재MM골병성골수억궤제중발휘일정작용.
Objective To culture osteoblast in vitro and evaluate CCL3 receptor CCR1 expression in patients with multiple myeloma (MM).Methods Bone marrow osteoblasts from MM patients were cultured in vitro with dexamethasone,β-sodium glycerophosphate and vitamin C,which were identified by alkaline phosphatase staining,Von Kossa' s staining.The CCL3 receptor expression was evaluated by flow cytometry.The morphology and quantity of osteoblast were observed after exposure to CCL3.Results Bone marrow osteoblasts from MM patients could be cultured in vitro and be identified by positive staining of alkaline phosphatase and Von Kossa' s.MM-derived osteoblasts expressed higher levels of CCR1 (74.48±7.31)％,compared with normal controls (48.35±8.81)％.Calcium deposition of osteoblasts after exposure to CCL3 was less than that of controls.Conclusions Bone marrow osteoblasts could be cultured in vitro from MM Patients.CCL3 may contribute to the development of myeloma bone disease.
