解剖学报
解剖學報
해부학보
ACTA ANATOMICA SINICA
2010年
1期
60-64
,共5页
马芹颖%顾平%王彦永%王铭维%崔冬生%温雅%刘力
馬芹穎%顧平%王彥永%王銘維%崔鼕生%溫雅%劉力
마근영%고평%왕언영%왕명유%최동생%온아%류력
骨髓基质细胞%神经干细胞%Shotgun蛋白质组学分析
骨髓基質細胞%神經榦細胞%Shotgun蛋白質組學分析
골수기질세포%신경간세포%Shotgun단백질조학분석
Bone marrow stromal cells%Neural stem cells%Shotgun proteomics
目的 通过Shotgun蛋白质组学分析,初步检测骨髓基质细胞(BMSCs)条件液中可能含有的蛋白质.方法 将BMSCs条件液混匀后经超滤浓缩分为大于5kD和小于5kD两部分,并培养神经干细胞(NSCs),观察其后代中神经元、星形胶质细胞和少突胶质细胞的比例,鉴定出能调节NSCs分化的部分,并进行Shotgun蛋白质组学分析.结果 BMSCs条件液分子量大于5kD的部分可以调节NSCs向神经元和少突胶质细胞方向分化,这部分条件液先经过聚丙烯酰胺凝胶电泳(SDS-PAGE)发现大部分蛋白集中在14kD以上,将蛋白条带酶解以后,经Shotgun分析共鉴定到456种蛋白质,其中154个相似蛋白质,17个假想蛋白质,56个未知蛋白质,剩余的229个蛋白质中大多为细胞骨架蛋白、分泌蛋白、信号转导蛋白、酶类和转运蛋白等.结论 BMSCs分泌的多种蛋白质对NSCs的分化起调节作用,明确了BMSCs条件液中可能含有的成分.
目的 通過Shotgun蛋白質組學分析,初步檢測骨髓基質細胞(BMSCs)條件液中可能含有的蛋白質.方法 將BMSCs條件液混勻後經超濾濃縮分為大于5kD和小于5kD兩部分,併培養神經榦細胞(NSCs),觀察其後代中神經元、星形膠質細胞和少突膠質細胞的比例,鑒定齣能調節NSCs分化的部分,併進行Shotgun蛋白質組學分析.結果 BMSCs條件液分子量大于5kD的部分可以調節NSCs嚮神經元和少突膠質細胞方嚮分化,這部分條件液先經過聚丙烯酰胺凝膠電泳(SDS-PAGE)髮現大部分蛋白集中在14kD以上,將蛋白條帶酶解以後,經Shotgun分析共鑒定到456種蛋白質,其中154箇相似蛋白質,17箇假想蛋白質,56箇未知蛋白質,剩餘的229箇蛋白質中大多為細胞骨架蛋白、分泌蛋白、信號轉導蛋白、酶類和轉運蛋白等.結論 BMSCs分泌的多種蛋白質對NSCs的分化起調節作用,明確瞭BMSCs條件液中可能含有的成分.
목적 통과Shotgun단백질조학분석,초보검측골수기질세포(BMSCs)조건액중가능함유적단백질.방법 장BMSCs조건액혼균후경초려농축분위대우5kD화소우5kD량부분,병배양신경간세포(NSCs),관찰기후대중신경원、성형효질세포화소돌효질세포적비례,감정출능조절NSCs분화적부분,병진행Shotgun단백질조학분석.결과 BMSCs조건액분자량대우5kD적부분가이조절NSCs향신경원화소돌효질세포방향분화,저부분조건액선경과취병희선알응효전영(SDS-PAGE)발현대부분단백집중재14kD이상,장단백조대매해이후,경Shotgun분석공감정도456충단백질,기중154개상사단백질,17개가상단백질,56개미지단백질,잉여적229개단백질중대다위세포골가단백、분비단백、신호전도단백、매류화전운단백등.결론 BMSCs분비적다충단백질대NSCs적분화기조절작용,명학료BMSCs조건액중가능함유적성분.
Objective Using shotgun mass spectrometry to detect proteins probably contained in bone marrow stromal cells(BMSCs) conditioned medium. Methods Mixed with BMSCs conditioned medium was divided into two parts which is(>5kD and <5kD) by means of ultrafiltration. The two parts were used to culture neural stem cells(NSCs) separately, and the proportions of neurons, astrocytes and oligodendrocytes in the offsprings of NSCs were calculated, then the effective part that could regulate the differentiation of NSCs was detected by Shotgun mass spectrometry. Results The BMSCs conditioned medium which is >5kD could promote the NSCs differentiate into more neurons and oligodendrocytes. The SDS-PAGE of this part showed that the most proteins were above 14kD, then the protein bands were enzymed. In total, 456 proteins were identified by Shotgun mass spectrometry after all the protein bands were enzymed, there were 154 similar proteins, 17 hypothetical proteins and 56 unknown proteins. And in the rest of 229 proteins, most of them were cytoskeletal proteins, secreted proteins, signal transduction proteins, enzymes, transporter and so on. Conclusion Many proteins secreted by BMSCs could regulate the differentiation of NSCs so as to prove the protein components probably existed in the BMSCs conditioned medium.
