中国组织工程研究
中國組織工程研究
중국조직공정연구
Journal of Clinical Rehabilitative Tissue Engineering Research
2012年
45期
8424-8429
,共6页
黎洪棉%柳大烈%赵培冉%梁双武
黎洪棉%柳大烈%趙培冉%樑雙武
려홍면%류대렬%조배염%량쌍무
人脂肪干细胞%自体富血小板血浆%组织工程%脂肪形成%血管发生%干细胞
人脂肪榦細胞%自體富血小闆血漿%組織工程%脂肪形成%血管髮生%榦細胞
인지방간세포%자체부혈소판혈장%조직공정%지방형성%혈관발생%간세포
背景:血运重建机制是组织工程化脂肪组织成功构建的决定性因素.
目的:观察脂肪干细胞与自体富血小板血浆和纤维蛋白胶载体复合物在体内构建血管化组织工程脂肪的可行性.
方法:从健康成年人吸脂术后的脂肪组织中分离脂肪干细胞并行原代及传代培养,将第3代经 BrdU 标记的脂肪干细胞向脂肪细胞定向诱导2周后,制成浓度为5×1010 L-1细胞悬液.由0.5 mL 细胞悬液、100μL 富血小板血浆工作液或 DMEM 培养基与0.5 mL 纤维蛋白胶组成实验组和对照组移植物,分别植入裸鼠背部皮下.
结果与结论:植入后8周取材:①实验组可见血管明显增生并长入材料,呈轻度纤维包裹;对照组有少量血管长入材料中,也有轻度纤维包裹现象.实验组新生组织湿质量大于对照组(P <0.01).②苏木精-伊红染色均可见移植物中有新生脂肪组织形成和不同程度的微血管长入;实验组微血管数多于对照组(P <0.01).③新生组织免疫荧光染色示,两组新生脂肪细胞的胞核及部分微血管内皮细胞的胞核呈现绿色荧光.说明脂肪干细胞与自体富血小板血浆和纤维蛋白胶载体复合物在体内可构建血管化组织工程脂肪,脂肪干细胞与自体富血小板血浆共同参与新生脂肪组织的血管化过程,自体富血小板血浆能促进组织工程构建物的血运重建,保证更多种子细胞的成活.
揹景:血運重建機製是組織工程化脂肪組織成功構建的決定性因素.
目的:觀察脂肪榦細胞與自體富血小闆血漿和纖維蛋白膠載體複閤物在體內構建血管化組織工程脂肪的可行性.
方法:從健康成年人吸脂術後的脂肪組織中分離脂肪榦細胞併行原代及傳代培養,將第3代經 BrdU 標記的脂肪榦細胞嚮脂肪細胞定嚮誘導2週後,製成濃度為5×1010 L-1細胞懸液.由0.5 mL 細胞懸液、100μL 富血小闆血漿工作液或 DMEM 培養基與0.5 mL 纖維蛋白膠組成實驗組和對照組移植物,分彆植入裸鼠揹部皮下.
結果與結論:植入後8週取材:①實驗組可見血管明顯增生併長入材料,呈輕度纖維包裹;對照組有少量血管長入材料中,也有輕度纖維包裹現象.實驗組新生組織濕質量大于對照組(P <0.01).②囌木精-伊紅染色均可見移植物中有新生脂肪組織形成和不同程度的微血管長入;實驗組微血管數多于對照組(P <0.01).③新生組織免疫熒光染色示,兩組新生脂肪細胞的胞覈及部分微血管內皮細胞的胞覈呈現綠色熒光.說明脂肪榦細胞與自體富血小闆血漿和纖維蛋白膠載體複閤物在體內可構建血管化組織工程脂肪,脂肪榦細胞與自體富血小闆血漿共同參與新生脂肪組織的血管化過程,自體富血小闆血漿能促進組織工程構建物的血運重建,保證更多種子細胞的成活.
배경:혈운중건궤제시조직공정화지방조직성공구건적결정성인소.
목적:관찰지방간세포여자체부혈소판혈장화섬유단백효재체복합물재체내구건혈관화조직공정지방적가행성.
방법:종건강성년인흡지술후적지방조직중분리지방간세포병행원대급전대배양,장제3대경 BrdU 표기적지방간세포향지방세포정향유도2주후,제성농도위5×1010 L-1세포현액.유0.5 mL 세포현액、100μL 부혈소판혈장공작액혹 DMEM 배양기여0.5 mL 섬유단백효조성실험조화대조조이식물,분별식입라서배부피하.
결과여결론:식입후8주취재:①실험조가견혈관명현증생병장입재료,정경도섬유포과;대조조유소량혈관장입재료중,야유경도섬유포과현상.실험조신생조직습질량대우대조조(P <0.01).②소목정-이홍염색균가견이식물중유신생지방조직형성화불동정도적미혈관장입;실험조미혈관수다우대조조(P <0.01).③신생조직면역형광염색시,량조신생지방세포적포핵급부분미혈관내피세포적포핵정현록색형광.설명지방간세포여자체부혈소판혈장화섬유단백효재체복합물재체내가구건혈관화조직공정지방,지방간세포여자체부혈소판혈장공동삼여신생지방조직적혈관화과정,자체부혈소판혈장능촉진조직공정구건물적혈운중건,보증경다충자세포적성활.
BACKGROUND: Revascularization mechanism is a determinal factor of successful construction of adipose tissue by tissue engineering.
@@@@OBJECTIVE: To investigate the feasibility of construction of vasuclarized adipose using adipose-derived stem cel s and autogeneic platelet-rich plasma carrier complex in vivo by tissue engineering.
@@@@METHODS: Adipose-derived stem cel s were isolated from the subcutaneous adipose tissue of healthy adult after liposuction, and primary culture and subculture of adipose-derived stem cel s were conducted. After being induced
@@@@towards adipocytes for 2 weeks, 5× 1010/L passage 3 cel suspension labeled by BrdU was prepared. Two groups were included: experimental group, in which 0.5 mL cel suspension, 100 μL platelet-rich plasma and 0.5 mL fibrin glue were implanted into the subcutaneous fascia of nude mice; control group in which 0.5 mL cel suspension, 100 μL DMEM and 0.5 mL fibrin glue were implanted into the subcutaneous fascia of nude mice.
@@@@RESULTS AND CONCLUSION: At 8 weeks after surgery, neogenetic vessels grew into the scaffolds and mild fiber encapsulation was observed in the experimental group, while few vessels grew into the scaffolds and mild fiber encapsulation was also observed in the control group. The wet weight of cambium in the experimental group was higher than that in the control group (P < 0.01). Hematoxylin-eosin staining showed the formation of neogenetic adipose tissues and the growth of micrangium in the implant. The number of micro vessels in the experimental group was greater than that in the control group (P < 0.01). The immunofluorescence staining of cambium showed that the cel nucleus of regenerated adipocytes and partial capil ary endothelium in both groups presented green fluorescence. It is feasible to prepare vasuclarized adipose using adipose-derived stem cel s and autogeneic platelet-rich plasma carrier complex in vivo by tissue engineering. Adipose-derived stem cel s and autogeneic platelet-rich plasma participate in neovascularization of neogenetic adipose tissue. The autogeneic platelet-rich plasma can promote revascularization of tissue-engineered complex and ensure survival of more seed cel s.