中华神经医学杂志
中華神經醫學雜誌
중화신경의학잡지
CHINESE JOURNAL OF NEUROMEDICINE
2009年
5期
468-471,475
,共5页
闫中杰%徐如祥%张洪钿%姜晓丹%林建浩%马旭%李西锋%赵信德
閆中傑%徐如祥%張洪鈿%薑曉丹%林建浩%馬旭%李西鋒%趙信德
염중걸%서여상%장홍전%강효단%림건호%마욱%리서봉%조신덕
雪旺细胞%细胞分化%骨髓基质细胞
雪旺細胞%細胞分化%骨髓基質細胞
설왕세포%세포분화%골수기질세포
Schwann cells%Cell differentiation%Bone marrow stromal cells
目的 研究骨髓基质细胞(BMSCs)在体外条件下向周围神经雪旰细胞(SCs)分化的可靠性. 方法 分离提取SD大鼠股骨和胫骨部位BMSCs.利用其贴壁生长的特性.培养纯化,传代扩增.用复合诱导因子(β.巯基乙醇+全反式维甲酸+血小板凝集抑制剂+m小板源性生长因子+碱性成纤维细胞生长因子)在体外诱导BMSCs分化.免疫细胞化学方法 检测P75、S-100及胶质原纤维酸性蛋白(GFAP)的表达,荧光实时定量PCR检测P75、S100及CD104的表达. 结果诱导后的BMSCs形态类似SCs,免疫荧光染色鉴定其具有SCs性质,表达SCs的表面标志物(GFAP、S100和P75).荧光实时定量PCR结果显示诱导后BMSCs S-100、CD104的表达量达到了SCs的表达量水平,但P75的表达量与SCs的表达量水平还有较大差距. 结论 体外诱导BMSCs可部分获得SCs的特征,传代后恢复至未诱导状态,这种预诱导加复合因子诱导的方法 尚待完善.
目的 研究骨髓基質細胞(BMSCs)在體外條件下嚮週圍神經雪旰細胞(SCs)分化的可靠性. 方法 分離提取SD大鼠股骨和脛骨部位BMSCs.利用其貼壁生長的特性.培養純化,傳代擴增.用複閤誘導因子(β.巰基乙醇+全反式維甲痠+血小闆凝集抑製劑+m小闆源性生長因子+堿性成纖維細胞生長因子)在體外誘導BMSCs分化.免疫細胞化學方法 檢測P75、S-100及膠質原纖維痠性蛋白(GFAP)的錶達,熒光實時定量PCR檢測P75、S100及CD104的錶達. 結果誘導後的BMSCs形態類似SCs,免疫熒光染色鑒定其具有SCs性質,錶達SCs的錶麵標誌物(GFAP、S100和P75).熒光實時定量PCR結果顯示誘導後BMSCs S-100、CD104的錶達量達到瞭SCs的錶達量水平,但P75的錶達量與SCs的錶達量水平還有較大差距. 結論 體外誘導BMSCs可部分穫得SCs的特徵,傳代後恢複至未誘導狀態,這種預誘導加複閤因子誘導的方法 尚待完善.
목적 연구골수기질세포(BMSCs)재체외조건하향주위신경설간세포(SCs)분화적가고성. 방법 분리제취SD대서고골화경골부위BMSCs.이용기첩벽생장적특성.배양순화,전대확증.용복합유도인자(β.구기을순+전반식유갑산+혈소판응집억제제+m소판원성생장인자+감성성섬유세포생장인자)재체외유도BMSCs분화.면역세포화학방법 검측P75、S-100급효질원섬유산성단백(GFAP)적표체,형광실시정량PCR검측P75、S100급CD104적표체. 결과유도후적BMSCs형태유사SCs,면역형광염색감정기구유SCs성질,표체SCs적표면표지물(GFAP、S100화P75).형광실시정량PCR결과현시유도후BMSCs S-100、CD104적표체량체도료SCs적표체량수평,단P75적표체량여SCs적표체량수평환유교대차거. 결론 체외유도BMSCs가부분획득SCs적특정,전대후회복지미유도상태,저충예유도가복합인자유도적방법 상대완선.
Objective To explore the reliability of acquiring Schwann cells from bone marrow stromal cells (BMSCs) by inducing their differentiation in vitro. Methods BMSCs isolated from the tibial and femural bone marrow of SD rats by adherent culture were passaged and induced with beta-mercaptoethanol followed by retinoic acid, forskolin, basic fibroblast growth factor (bFGF), platelet-derived growth factor (PDGF) and heregulin. Immunocytochemistry was performed to identify the expression of the Schwann cell markers P75, S100 and glial fibrillary acidic portein (GFAP), and fluorescent real-time quantitative RT-PCR was used to detect the mRNA expressions of P75, S100 and CD104 in the induced BMSCs. Results The induced BMSCs exhibited morphological changes into cells resembling primary cultured Schwann ceils, and expressed P75, S100 and GFAP proteins, as shown by immunofluorescent staining, at the levels similar to those in Schwann cells. The induced cells, however, remained negative for P75 mRNA expression as demonstrated by RT-PCR. Conclusion The BMSCs can be induced to partially acquire the phenotypic features of Schwann cells, suggesting the strong plasticity of the BMSCs. The protocol of pre-induction followed by induction with an array of agents still needs further modification to induce the in vitro differentiation of BMSCs into Schwann-like cells.
