中国组织工程研究
中國組織工程研究
중국조직공정연구
Journal of Clinical Rehabilitative Tissue Engineering Research
2013年
20期
3611-3617
,共7页
王正东%颜南%潘峰%杨芳莉%刘自力%姜海波%臧晋%牟军%柏树令
王正東%顏南%潘峰%楊芳莉%劉自力%薑海波%臧晉%牟軍%柏樹令
왕정동%안남%반봉%양방리%류자력%강해파%장진%모군%백수령
组织构建%骨组织构建%破骨细胞%培养%诱导%鉴定%噬骨%1,25(OH)2D3%破骨样细胞%单核细胞%Wistar大鼠
組織構建%骨組織構建%破骨細胞%培養%誘導%鑒定%噬骨%1,25(OH)2D3%破骨樣細胞%單覈細胞%Wistar大鼠
조직구건%골조직구건%파골세포%배양%유도%감정%서골%1,25(OH)2D3%파골양세포%단핵세포%Wistar대서
tissue construction%bone tissue construction%osteoclasts%culture%induction%identification%bone absorption%1,25(OH) 2 D 3%osteoclast-like cells%mononuclear cells%Wistar rats
背景:原代培养的破骨细胞数量少,而诱导培养产生的破骨样细胞数量多,能够满足一些研究骨代谢实验的要求.但是两种细胞在特异酶及噬骨能力方面是否具有相同的效果,到目前为止,还没有详尽的数据来支持.目的:比较大鼠原代分离的破骨细胞及诱导形成的破骨样细胞噬骨能力上的差异.方法:取24 h内新生Wistar大鼠四肢长骨,酶消化法分离培养破骨细胞,将破骨细胞培养过程中消化下来的骨髓单核细胞加入1,25(OH)2 D 3结果与结论:诱导第9天,破骨样细胞数目为破骨细胞数目的11倍,其形态与原代消化获得的细胞形态相同.抗酒石酸酸性磷酸酶染色呈阳性.甲苯胺蓝染色显示两组破骨细胞在骨片上培养时产生骨陷凹面积及深度差异无显著性意义.结果显示破骨样细胞的形态、特异酶及噬骨能力与破骨细胞无差异.诱导生成破骨样细胞.苏木精-伊红染色、抗酒石酸酸性磷酸酶(TRAP)染色鉴定.用甲苯胺蓝染色共培养骨片比较两组破骨细胞噬骨能力.
揹景:原代培養的破骨細胞數量少,而誘導培養產生的破骨樣細胞數量多,能夠滿足一些研究骨代謝實驗的要求.但是兩種細胞在特異酶及噬骨能力方麵是否具有相同的效果,到目前為止,還沒有詳儘的數據來支持.目的:比較大鼠原代分離的破骨細胞及誘導形成的破骨樣細胞噬骨能力上的差異.方法:取24 h內新生Wistar大鼠四肢長骨,酶消化法分離培養破骨細胞,將破骨細胞培養過程中消化下來的骨髓單覈細胞加入1,25(OH)2 D 3結果與結論:誘導第9天,破骨樣細胞數目為破骨細胞數目的11倍,其形態與原代消化穫得的細胞形態相同.抗酒石痠痠性燐痠酶染色呈暘性.甲苯胺藍染色顯示兩組破骨細胞在骨片上培養時產生骨陷凹麵積及深度差異無顯著性意義.結果顯示破骨樣細胞的形態、特異酶及噬骨能力與破骨細胞無差異.誘導生成破骨樣細胞.囌木精-伊紅染色、抗酒石痠痠性燐痠酶(TRAP)染色鑒定.用甲苯胺藍染色共培養骨片比較兩組破骨細胞噬骨能力.
배경:원대배양적파골세포수량소,이유도배양산생적파골양세포수량다,능구만족일사연구골대사실험적요구.단시량충세포재특이매급서골능력방면시부구유상동적효과,도목전위지,환몰유상진적수거래지지.목적:비교대서원대분리적파골세포급유도형성적파골양세포서골능력상적차이.방법:취24 h내신생Wistar대서사지장골,매소화법분리배양파골세포,장파골세포배양과정중소화하래적골수단핵세포가입1,25(OH)2 D 3결과여결론:유도제9천,파골양세포수목위파골세포수목적11배,기형태여원대소화획득적세포형태상동.항주석산산성린산매염색정양성.갑분알람염색현시량조파골세포재골편상배양시산생골함요면적급심도차이무현저성의의.결과현시파골양세포적형태、특이매급서골능력여파골세포무차이.유도생성파골양세포.소목정-이홍염색、항주석산산성린산매(TRAP)염색감정.용갑분알람염색공배양골편비교량조파골세포서골능력.
@@@@BACKGROUND: There are less primary cultured osteoclasts, but a large number of osteoclast-like cells produced by induced culture, which is able to meet the requirements of some experimental studies of bone metabolism. But there are no detailed data to identify whether the osteoclasts and osteoclast-like cells have the similar effect on specific enzymes and bone absorption capacity. OBJECTIVE: To compare the difference of bone absorption ability between the primary cultured osteoclasts and induce cultured osteoclast-like cells from the rats. METHODS: The long bones were obtained from the limbs of 24 hours newborn Wistar rates, and the enzyme digestion was used to isolate and culture osteoclasts; then bone marrow mononuclear cell s separated from the osteoclasts during culture were added with 1,25(OH) 2 D 3 RESULTS AND CONCLUSION: The number of the osteoclast-like cell s was 11 times that of osteoclasts at 9 days after induction. The morphology of osteoclast-like cell s was similar with that of the primary cultured osteoclasts. The osteoclast-like cell s were positive for tartrate-resistant acid phosphatase. Toluidine blue staining results showed there was no significant difference in bone lacuna area and depth produced by the osteoclasts cultured on the bone slices between two groups. These findings show that there are no significant differences in morphology, specific enzymes and bone absorption capacity between osteoclast-like cell s and osteoclasts. to induce to generate the osteoclast-like cell s. The cell s were identified with heamtoxylin-eosin staining and tartrate-resistant acid phosphatase staining. The bone absorption ability of the osteoclasts in two groups was compared with toluidine blue staining.