牙体牙髓牙周病学杂志
牙體牙髓牙週病學雜誌
아체아수아주병학잡지
CHINESE JOURNAL OF CONSERVATIVE DENTISTRY
2014年
6期
331-337
,共7页
郑桂婷%徐燕%赵璇%沈继龙
鄭桂婷%徐燕%趙璇%瀋繼龍
정계정%서연%조선%침계룡
根端复合体%脂肪干细胞%条件培养液%增殖%分化
根耑複閤體%脂肪榦細胞%條件培養液%增殖%分化
근단복합체%지방간세포%조건배양액%증식%분화
developing apical complex (DAC)%adipose tissue-derived stem cells (ADSCs)%conditioned medi-um%proliferation%differentiation
目的：初步探讨发育期根端复合体（developing apical complex，DAC）条件培养基（DAC condi-tioned medium，DACCM）对脂肪干细胞（adipose tissue-derived stem cells，ADSCs）增殖和分化的影响。方法：取出生20 d SD大鼠分离其下颌磨牙DAC，并采用组织块联合酶消化法培养DAC细胞；经HE、免疫组化染色鉴定DAC后，收集原代培养的DAC细胞上清，分别用过滤和不过滤的方式制备不同条件培养基并用其对ADSCs进行培养；分别于培养3、5、7 d 各时间点，MTT法检测ADSCs增殖能力；用碱性磷酸酶（ALP）试剂盒及反转录酶-聚合酶链反应（RT-PCR）检测ADSCs的ALP蛋白活性及其基因的表达水平。结果：免疫组化染色显示，培养的DAC细胞CK-14、vimentin均为阳性表达；DACCM-未滤培养组ADSCs的增殖活性、ALP蛋白活性以及ALP基因表达水平均高于单纯α-MEM培养组和 DACCM-过滤培养组，差异均有统计学意义（P＜0．05）。结论：不经过滤的DAC细胞上清制备的条件培养基对ADSCs增殖、分化的促进作用更明显。
目的：初步探討髮育期根耑複閤體（developing apical complex，DAC）條件培養基（DAC condi-tioned medium，DACCM）對脂肪榦細胞（adipose tissue-derived stem cells，ADSCs）增殖和分化的影響。方法：取齣生20 d SD大鼠分離其下頜磨牙DAC，併採用組織塊聯閤酶消化法培養DAC細胞；經HE、免疫組化染色鑒定DAC後，收集原代培養的DAC細胞上清，分彆用過濾和不過濾的方式製備不同條件培養基併用其對ADSCs進行培養；分彆于培養3、5、7 d 各時間點，MTT法檢測ADSCs增殖能力；用堿性燐痠酶（ALP）試劑盒及反轉錄酶-聚閤酶鏈反應（RT-PCR）檢測ADSCs的ALP蛋白活性及其基因的錶達水平。結果：免疫組化染色顯示，培養的DAC細胞CK-14、vimentin均為暘性錶達；DACCM-未濾培養組ADSCs的增殖活性、ALP蛋白活性以及ALP基因錶達水平均高于單純α-MEM培養組和 DACCM-過濾培養組，差異均有統計學意義（P＜0．05）。結論：不經過濾的DAC細胞上清製備的條件培養基對ADSCs增殖、分化的促進作用更明顯。
목적：초보탐토발육기근단복합체（developing apical complex，DAC）조건배양기（DAC condi-tioned medium，DACCM）대지방간세포（adipose tissue-derived stem cells，ADSCs）증식화분화적영향。방법：취출생20 d SD대서분리기하합마아DAC，병채용조직괴연합매소화법배양DAC세포；경HE、면역조화염색감정DAC후，수집원대배양적DAC세포상청，분별용과려화불과려적방식제비불동조건배양기병용기대ADSCs진행배양；분별우배양3、5、7 d 각시간점，MTT법검측ADSCs증식능력；용감성린산매（ALP）시제합급반전록매-취합매련반응（RT-PCR）검측ADSCs적ALP단백활성급기기인적표체수평。결과：면역조화염색현시，배양적DAC세포CK-14、vimentin균위양성표체；DACCM-미려배양조ADSCs적증식활성、ALP단백활성이급ALP기인표체수평균고우단순α-MEM배양조화 DACCM-과려배양조，차이균유통계학의의（P＜0．05）。결론：불경과려적DAC세포상청제비적조건배양기대ADSCs증식、분화적촉진작용경명현。
AIM:To study the effects of the developing apical complex (DAC)cells conditioned medium (DACCM)on the proliferation and differentiation of adipose tissue-derived stem cells (ADSCs).METHODS:DAC cells,separated from the 20 d postnatal SD rat,were cultured by tissue block combined with enzyme digestion method. DACs was identified by HE staining and immumohistochemical staining.DACCM was collected and prepared by filtra-tion and none-filtration (DACCM-F and DACCM-UF)respectively.The proliferation of ADSCs was detected by MTT assay.The mRNA and protein expression of alkaline phosphatase (ALP)were detected by RT-PCR and ALP detection Kit respectively.SPASS13.0 was used for data analysis.RESULTS:DAC cells were obtained from the developing roots of the SD rat.The cells were CK-14 and vimentin positive.The proliferation,ALP mRNA and protein expression of ADSCs cultured in DACCM-UF exceeded those of the cells cultured in α-MEM or DACCM-F(P<0.05 )CON-CLUSION:DACCM-UF may stimulate the proliferation and differentiation of ADSCs.
