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
10期
1591-1596
,共6页
白金萍%李秀英%李雪%杨麒巍%刘颖男%王轶敏
白金萍%李秀英%李雪%楊麒巍%劉穎男%王軼敏
백금평%리수영%리설%양기외%류영남%왕질민
干细胞%间充质干细胞%胎盘间充质干细胞%胎盘%增殖能力
榦細胞%間充質榦細胞%胎盤間充質榦細胞%胎盤%增殖能力
간세포%간충질간세포%태반간충질간세포%태반%증식능력
stem cells%mesenchymal stem cells%placenta%cell proliferation
背景：胎盘间充质干细胞的增殖能力比骨髓间充质干细胞增殖能力强，但是否每个代次胎盘间充质干细胞的增殖能力都相同尚无定论。
<br> 目的：分析不同代次胎盘间充质干细胞的增殖能力。
<br> 方法：采用酶消化法分离人胎盘间充质干细胞，并利用流式细胞仪检测干细胞表面标志物，并进行成脂成骨诱导分化。采用细胞计数和BrdU方法检测细胞的增殖能力，根据倍增时间公式计算倍增时间，并进行长期传代。
<br> 结果与结论：胎盘间充质干细胞阳性表达CD44、CD90和CD105，阴性表达CD14、CD34和CD45；成脂诱导21 d后油红O染色镜下可见红色脂肪滴，成骨诱导35 d茜素红染色镜下可见钙盐结节。细胞从第1代开始计数，直到第15代共获得3.5×1011个细胞。P5代比P10和P15代胎盘间充质干细胞的增殖能力强，倍增时间短。长期传代，胎盘间充质干细胞传代到第25代时，其中3例标本细胞形态完全发生改变，呈铺路石样；另外2例标本几乎无贴壁细胞。提示胎盘间充质干细胞可以长期传代到第25代，不同代次的胎盘间充质干细胞增殖能力不同，P5代增殖能力明显高于P10和P15代，临床应用应以P5代之前为宜。
揹景：胎盤間充質榦細胞的增殖能力比骨髓間充質榦細胞增殖能力彊，但是否每箇代次胎盤間充質榦細胞的增殖能力都相同尚無定論。
<br> 目的：分析不同代次胎盤間充質榦細胞的增殖能力。
<br> 方法：採用酶消化法分離人胎盤間充質榦細胞，併利用流式細胞儀檢測榦細胞錶麵標誌物，併進行成脂成骨誘導分化。採用細胞計數和BrdU方法檢測細胞的增殖能力，根據倍增時間公式計算倍增時間，併進行長期傳代。
<br> 結果與結論：胎盤間充質榦細胞暘性錶達CD44、CD90和CD105，陰性錶達CD14、CD34和CD45；成脂誘導21 d後油紅O染色鏡下可見紅色脂肪滴，成骨誘導35 d茜素紅染色鏡下可見鈣鹽結節。細胞從第1代開始計數，直到第15代共穫得3.5×1011箇細胞。P5代比P10和P15代胎盤間充質榦細胞的增殖能力彊，倍增時間短。長期傳代，胎盤間充質榦細胞傳代到第25代時，其中3例標本細胞形態完全髮生改變，呈鋪路石樣；另外2例標本幾乎無貼壁細胞。提示胎盤間充質榦細胞可以長期傳代到第25代，不同代次的胎盤間充質榦細胞增殖能力不同，P5代增殖能力明顯高于P10和P15代，臨床應用應以P5代之前為宜。
배경：태반간충질간세포적증식능력비골수간충질간세포증식능력강，단시부매개대차태반간충질간세포적증식능력도상동상무정론。
<br> 목적：분석불동대차태반간충질간세포적증식능력。
<br> 방법：채용매소화법분리인태반간충질간세포，병이용류식세포의검측간세포표면표지물，병진행성지성골유도분화。채용세포계수화BrdU방법검측세포적증식능력，근거배증시간공식계산배증시간，병진행장기전대。
<br> 결과여결론：태반간충질간세포양성표체CD44、CD90화CD105，음성표체CD14、CD34화CD45；성지유도21 d후유홍O염색경하가견홍색지방적，성골유도35 d천소홍염색경하가견개염결절。세포종제1대개시계수，직도제15대공획득3.5×1011개세포。P5대비P10화P15대태반간충질간세포적증식능력강，배증시간단。장기전대，태반간충질간세포전대도제25대시，기중3례표본세포형태완전발생개변，정포로석양；령외2례표본궤호무첩벽세포。제시태반간충질간세포가이장기전대도제25대，불동대차적태반간충질간세포증식능력불동，P5대증식능력명현고우P10화P15대，림상응용응이P5대지전위의。
BACKGROUND:The mesenchymal stem cells which are from the placenta have better proliferative potential than the bone marrow mesenchymal stem cells. It is inconclusive, however, whether placenta-derived mesenchymal stem cells at different passages have the same proliferative ability.
<br> OBJECTIVE:To analyze the proliferative ability of placenta-derived mesenchymal stem cells.
<br> METHODS:Placenta-derived human mesenchymal stem cells were isolated with enzyme digestion, and flow cytometry was used to detect the cellsurface markers. After osteogenic and adipogenic induction, cellproliferation detection was carried out using cellcounting and BrdU analysis. celldoubling time was calculated based on formula. The maximum cellpassage was determined by long term cellculture until the celldied or differentiated.
<br> RESULTS AND CONCLUSION:CD44, CD90 and CD105 were highly expressed in placenta-derived mesenchymal stem cells, while CD14, CD34 and CD45 were negatively expressed. After 21 days of adipogenic induction, oil red O staining showed red fat droplets;after 35 days of osteogenic induction, alizarin red staining showed calcium nodules. cellcounting started from passage 1 to 25, and cellnumber reached to 3.5×1011 at passage 15. cellproliferation at passage 5 was stronger with shorter doubling time than that at passages 10 and 15. When cells reached to passage 25, three specimens completely altered their cellmorphology with cobblestone structure, and another two specimens lost adherent features. These findings indicate that placenta-derived mesenchymal stem cells can be long-term subcultured upon to passage 25. Different proliferation capacities are found in different passages of placenta-derived mesenchymal stem cells. The proliferation ability of passage 5 placenta-derived mesenchymal stem cells is stronger than that of passage 10 and 15 cells. Placenta-derived mesenchymal stem cells at passages 0-5 are the important source for stem celltherapy and tissue engineering.
