中华实验外科杂志
中華實驗外科雜誌
중화실험외과잡지
CHINESE JOURNAL OF EXPERIMENTAL SURGERY
2012年
6期
1085-1088
,共4页
察鹏飞%高建华%陈阳%鲁峰
察鵬飛%高建華%陳暘%魯峰
찰붕비%고건화%진양%로봉
脂肪来源干细胞%细胞外基质%支架%组织工程
脂肪來源榦細胞%細胞外基質%支架%組織工程
지방래원간세포%세포외기질%지가%조직공정
Adipose-derived stem cells%Extracellular matrix%Scaffold%Tissue engineering
目的 探讨人脂肪组织细胞外基质(ECM)支架联合人脂肪来源干细胞(ADSCs)构建工程化脂肪组织的可行性.方法 以酶消化法从人抽脂术抽吸物脂质部分获取人ADSCs,体外进行多向分化诱导鉴定,并行DiI荧光标记.从抽脂术的脂质部分分离提取人脂肪组织细胞外基质,经过低温冻干、粉碎、灭菌等处理,制备成粉末状,电镜扫描观察表面特征并将其与ADSCs进行黏附实验,探讨其作为支架材料的可行性.收集人ADSCs,以2×109/L的细胞密度与提取的细胞外基质支架复合后移植于裸鼠背部皮下,同鼠对侧背部皮下移植ECM支架和细胞培养液作为对照,每侧移植0.5 ml,共6只实验鼠.8周后取材,称量标本湿重.取出的标本行苏木素-伊红(HE)染色和油红O染色进行定性判断,分析人脂肪组织ECM支架联合人ADSCs构建工程化脂肪组织的能力.结果 从脂肪组织中分离得到人ADSCs和ECM支架.ADSCs在相应的诱导环境下能够分化成为脂肪细胞、骨细胞和软骨细胞.ECM支架电镜扫描和大体观察具有疏松、多孔的结构特征,适合ADSCs的黏附生长.ADSCs与支架相容性良好,黏附率达(89.87±2.59)%,细胞在支架表面可充分伸展生长.体内移植8周后,实验组和对照组都能够形成新生物,湿重比较实验组较对照组重(P<0.05).经HE切片及油红O染色均证实实验组形成成熟的脂肪组织,对照组不能形成脂肪组织.结论 人脂肪组织ECM支架联合人ADSCs在体内能够成功构建成熟的脂肪组织,8周后支架并无明显吸收.
目的 探討人脂肪組織細胞外基質(ECM)支架聯閤人脂肪來源榦細胞(ADSCs)構建工程化脂肪組織的可行性.方法 以酶消化法從人抽脂術抽吸物脂質部分穫取人ADSCs,體外進行多嚮分化誘導鑒定,併行DiI熒光標記.從抽脂術的脂質部分分離提取人脂肪組織細胞外基質,經過低溫凍榦、粉碎、滅菌等處理,製備成粉末狀,電鏡掃描觀察錶麵特徵併將其與ADSCs進行黏附實驗,探討其作為支架材料的可行性.收集人ADSCs,以2×109/L的細胞密度與提取的細胞外基質支架複閤後移植于裸鼠揹部皮下,同鼠對側揹部皮下移植ECM支架和細胞培養液作為對照,每側移植0.5 ml,共6隻實驗鼠.8週後取材,稱量標本濕重.取齣的標本行囌木素-伊紅(HE)染色和油紅O染色進行定性判斷,分析人脂肪組織ECM支架聯閤人ADSCs構建工程化脂肪組織的能力.結果 從脂肪組織中分離得到人ADSCs和ECM支架.ADSCs在相應的誘導環境下能夠分化成為脂肪細胞、骨細胞和軟骨細胞.ECM支架電鏡掃描和大體觀察具有疏鬆、多孔的結構特徵,適閤ADSCs的黏附生長.ADSCs與支架相容性良好,黏附率達(89.87±2.59)%,細胞在支架錶麵可充分伸展生長.體內移植8週後,實驗組和對照組都能夠形成新生物,濕重比較實驗組較對照組重(P<0.05).經HE切片及油紅O染色均證實實驗組形成成熟的脂肪組織,對照組不能形成脂肪組織.結論 人脂肪組織ECM支架聯閤人ADSCs在體內能夠成功構建成熟的脂肪組織,8週後支架併無明顯吸收.
목적 탐토인지방조직세포외기질(ECM)지가연합인지방래원간세포(ADSCs)구건공정화지방조직적가행성.방법 이매소화법종인추지술추흡물지질부분획취인ADSCs,체외진행다향분화유도감정,병행DiI형광표기.종추지술적지질부분분리제취인지방조직세포외기질,경과저온동간、분쇄、멸균등처리,제비성분말상,전경소묘관찰표면특정병장기여ADSCs진행점부실험,탐토기작위지가재료적가행성.수집인ADSCs,이2×109/L적세포밀도여제취적세포외기질지가복합후이식우라서배부피하,동서대측배부피하이식ECM지가화세포배양액작위대조,매측이식0.5 ml,공6지실험서.8주후취재,칭량표본습중.취출적표본행소목소-이홍(HE)염색화유홍O염색진행정성판단,분석인지방조직ECM지가연합인ADSCs구건공정화지방조직적능력.결과 종지방조직중분리득도인ADSCs화ECM지가.ADSCs재상응적유도배경하능구분화성위지방세포、골세포화연골세포.ECM지가전경소묘화대체관찰구유소송、다공적결구특정,괄합ADSCs적점부생장.ADSCs여지가상용성량호,점부솔체(89.87±2.59)%,세포재지가표면가충분신전생장.체내이식8주후,실험조화대조조도능구형성신생물,습중비교실험조교대조조중(P<0.05).경HE절편급유홍O염색균증실실험조형성성숙적지방조직,대조조불능형성지방조직.결론 인지방조직ECM지가연합인ADSCs재체내능구성공구건성숙적지방조직,8주후지가병무명현흡수.
Objective To explore the possibility of building tissue-engineered adipose tissue with human adipose-derived stem cells (ADSCs) and adipose tissue extracellular matrix (ECM) scaffold,and provide experimental basis for clinical application of tissue-engineered adipose tissue for the repair of soft tissue defects.Methods ADSCs were isolated from adipose tissue by liposuction with the method of enzymatic digestion.The cells were induced for three-line differentiation and marked with DiI.Human ECM was extracted from adipose tissue obtained by liposuction.The ECM was freeze-dried,sterilized and crushed to powders.The surface characteristics of ECM were observed under the electron microscopy in vitro.The ADSCs were adhered to ECM powers,and the feasibility of ECM powers as scaffold was explorted.Human ADSCs in a density of 2 × 109/L were collected,and mixed them with ECM powers.They were transplanted subcutaneously into nude mice,and the opposite side of the same mice transplanted with scaffold only served as control group.There were 6 mice in total,and each side was transplanted by 0.5 ml.At 8th week post-transplantation,the mice were sacrificed and the implanted tissue were taken to record the wet weight,The hematoxylin and eosin (HE) sections and red oil O staining were detected.Two-sample test of SPSS 13.0 was used for statistical comparison between wet-determination.Results The ADSCs and human ECM powder were obtained successfully from adipose tissue.The ADSCs could differentiate into adipose cells,bone cells and chondrocytes.Scanning election microscopy (SEM) images showed that the powers had porous structure to which ADSCs could adhere easily,with the adhesion rate being (89.87 ±2.59)%.Eight weeks after transplantation,both experimental and control groups formed new tissues.There were significant differences between two groups ( P < 0.05 ) in the wet weight.HE and red oil O staining confirmed that the experimental group could form mature adipose tissue.Conclusion ADSCs and human ECM scaffold could form tissue-engineered adipose tissuein vivo.It can be used as an ideal method to construct tissue-engineered adipose tissue.