中国组织工程研究
中國組織工程研究
중국조직공정연구
Journal of Clinical Rehabilitative Tissue Engineering Research
2014年
6期
835-840
,共6页
邹斌%宗少晖%曾高峰%方晔%高太行
鄒斌%宗少暉%曾高峰%方曄%高太行
추빈%종소휘%증고봉%방엽%고태행
干细胞%骨髓干细胞%骨髓间充质干细胞%α-玉米赤霉醇%成骨细胞分化%碱性磷酸酶%骨钙素
榦細胞%骨髓榦細胞%骨髓間充質榦細胞%α-玉米赤黴醇%成骨細胞分化%堿性燐痠酶%骨鈣素
간세포%골수간세포%골수간충질간세포%α-옥미적매순%성골세포분화%감성린산매%골개소
bone marrow%mesenchymal stem cells%phytoestrogens%celldifferentiation%alkaline phosphatase%osteocalcin
背景：骨髓间充质干细胞具有向成骨细胞分化的能力，植物雌激素α-玉米赤霉醇对小鼠骨髓间充质干细胞的成骨分化可能有促进作用。
<br> 目的：探讨α-玉米赤霉醇对小鼠骨髓间充质干细胞向成骨细胞分化的影响。
<br> 方法：采用全骨髓培养差速贴壁法体外分离培养小鼠骨髓间充质干细胞，取第3代细胞分为6组：非诱导组加含体积分数为10%胎牛血清、1%双抗的 IMDM 普通培养基；空白诱导组仅加入等量成骨条件培养基(含0.1μmol/L 地塞米松、10 mmol/Lβ-甘油磷酸钠、50μmol/L 维生素C)；阳性对照诱导组加入等量成骨条件培养基及10-8 mol/L雌二醇；高、中、低α-玉米赤霉醇组分别加入成骨条件培养基及10-5，10-6，10-7 mol/L梯度浓度的α-玉米赤霉醇。倒置显微镜下观察细胞形态变化，MTT 实验测定细胞增殖状态绘制细胞生长曲线，酶联免疫吸附法测定碱性磷酸酶、骨钙素的表达。
<br> 结果与结论：非诱导组细胞呈长梭形，生长密集时有接触抑制，各诱导组的细胞增殖受促进，诱导后细胞呈三角形、多角形、不规则形状，细胞可重叠生长；非诱导组低表达碱性磷酸酶、骨钙素，低浓度(10-7 mol/L)α-玉米赤霉醇组和阳性对照诱导组高表达碱性磷酸酶、骨钙素，与空白诱导组相比，差异有非常显著性意义(P<0.01)。结果说明α-玉米赤霉醇可以促进骨髓间充质干细胞的增殖，低浓度α-玉米赤霉醇可显著促进体外培养的小鼠骨髓间充质干细胞成骨分化。而上调碱性磷酸酶、骨钙素的表达量可能是α-玉米赤霉醇诱导促进骨髓间充质干细胞骨向分化的作用机制之一。
揹景：骨髓間充質榦細胞具有嚮成骨細胞分化的能力，植物雌激素α-玉米赤黴醇對小鼠骨髓間充質榦細胞的成骨分化可能有促進作用。
<br> 目的：探討α-玉米赤黴醇對小鼠骨髓間充質榦細胞嚮成骨細胞分化的影響。
<br> 方法：採用全骨髓培養差速貼壁法體外分離培養小鼠骨髓間充質榦細胞，取第3代細胞分為6組：非誘導組加含體積分數為10%胎牛血清、1%雙抗的 IMDM 普通培養基；空白誘導組僅加入等量成骨條件培養基(含0.1μmol/L 地塞米鬆、10 mmol/Lβ-甘油燐痠鈉、50μmol/L 維生素C)；暘性對照誘導組加入等量成骨條件培養基及10-8 mol/L雌二醇；高、中、低α-玉米赤黴醇組分彆加入成骨條件培養基及10-5，10-6，10-7 mol/L梯度濃度的α-玉米赤黴醇。倒置顯微鏡下觀察細胞形態變化，MTT 實驗測定細胞增殖狀態繪製細胞生長麯線，酶聯免疫吸附法測定堿性燐痠酶、骨鈣素的錶達。
<br> 結果與結論：非誘導組細胞呈長梭形，生長密集時有接觸抑製，各誘導組的細胞增殖受促進，誘導後細胞呈三角形、多角形、不規則形狀，細胞可重疊生長；非誘導組低錶達堿性燐痠酶、骨鈣素，低濃度(10-7 mol/L)α-玉米赤黴醇組和暘性對照誘導組高錶達堿性燐痠酶、骨鈣素，與空白誘導組相比，差異有非常顯著性意義(P<0.01)。結果說明α-玉米赤黴醇可以促進骨髓間充質榦細胞的增殖，低濃度α-玉米赤黴醇可顯著促進體外培養的小鼠骨髓間充質榦細胞成骨分化。而上調堿性燐痠酶、骨鈣素的錶達量可能是α-玉米赤黴醇誘導促進骨髓間充質榦細胞骨嚮分化的作用機製之一。
배경：골수간충질간세포구유향성골세포분화적능력，식물자격소α-옥미적매순대소서골수간충질간세포적성골분화가능유촉진작용。
<br> 목적：탐토α-옥미적매순대소서골수간충질간세포향성골세포분화적영향。
<br> 방법：채용전골수배양차속첩벽법체외분리배양소서골수간충질간세포，취제3대세포분위6조：비유도조가함체적분수위10%태우혈청、1%쌍항적 IMDM 보통배양기；공백유도조부가입등량성골조건배양기(함0.1μmol/L 지새미송、10 mmol/Lβ-감유린산납、50μmol/L 유생소C)；양성대조유도조가입등량성골조건배양기급10-8 mol/L자이순；고、중、저α-옥미적매순조분별가입성골조건배양기급10-5，10-6，10-7 mol/L제도농도적α-옥미적매순。도치현미경하관찰세포형태변화，MTT 실험측정세포증식상태회제세포생장곡선，매련면역흡부법측정감성린산매、골개소적표체。
<br> 결과여결론：비유도조세포정장사형，생장밀집시유접촉억제，각유도조적세포증식수촉진，유도후세포정삼각형、다각형、불규칙형상，세포가중첩생장；비유도조저표체감성린산매、골개소，저농도(10-7 mol/L)α-옥미적매순조화양성대조유도조고표체감성린산매、골개소，여공백유도조상비，차이유비상현저성의의(P<0.01)。결과설명α-옥미적매순가이촉진골수간충질간세포적증식，저농도α-옥미적매순가현저촉진체외배양적소서골수간충질간세포성골분화。이상조감성린산매、골개소적표체량가능시α-옥미적매순유도촉진골수간충질간세포골향분화적작용궤제지일。
BACKGROUND:Bone marrow mesenchymal stem cells can differentiate into osteoblasts, andα-zearalanol can promote the osteogenic differentiation of mouse bone marrow mesenchymal stem cells.
<br> OBJECTIVE:To observe the effects ofα-zearalanol on the osteogenic differentiation of mouse bone marrow mesenchymal stem cells.
<br> METHODS:Mouse bone marrow mesenchymal stem cells were isolated through whole bone marrow adherence method. Passage 3 cells were divided into non-induced group (cultured by the Iscove’s modified Dulbecco’s medium containing 10%fetal bovine serum and 1%penicil in and streptomycin), blank induced group (induced by the classic osteoblast-induced system with 0.1μmol/L dexamethasone, 10 mmol/Lβ-glycerine sodium, and 50μmol/L vitamin C), positive control induced group (induced by the classic osteoblast-induced system and 10-8 mol/L oestrogen), and low-, median-, high-concentrationα-zearalanol groups (induced by the classic osteoblast-induced system and 10-7, 10-6, 10-5 mol/Lα-zearalanol). The morphological changes of induced cells were observed by phase contrast microscopy, and the growth curves of cultured cells were plotted based on MTT results. The expression of alkaline phosphatase and osteocalcin were detected by ELISA.
<br> RESULTS AND CONCLUSION:In the non-induced group, cells showed long fusiform shape mostly, and contact inhibition was observed while the cells were close-packed. After induced, cells proliferated effectively, showed triangle, astero-form, polygon or irregular shape, and clumps and multilayer of cells became evident. Compared to both the blank induced group, the alkaline phosphatase activity and osteocalcin expression were significantly lower in the non-induced group, but higher in the positive control induced group and low concentrationα-zearalanol group (P<0.01). These findings indicate thatα-zearalanol can promote the proliferation of bone marrow mesenchymal stem cells, and enhance the expression of alkaline phosphatase and osteocalcin expression, especial y in the low concentration group. This demonstrates that low concentrationα-zearalanol obviously enhances the osteogenic differentiation of mouse bone marrow mesenchymal stem cells in vitro. Up-regulating the release of alkaline phosphatase and osteocalcin may be one of the mechanisms by whichα-zearalanol promotes osteogenic differentiation of bone marrow mesenchymal stem cells.
