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自体脂肪干细胞复合胶原海绵多孔支架构建大鼠乳腺脂肪组织的初步研究

Adipose tissue engineering of the breast with adipose-derived stem cells and collagen sponge scaffolds in rats

摘要目的 探讨自体脂肪干细胞复合胶原海绵多孔支架构建大鼠乳腺脂肪组织的可行性.方法 采用胶原酶消化、离心方法分离并培养6只雌性SD大鼠的脂肪干细胞.对脂肪干细胞进行成骨、成脂诱导及鉴定.将自体脂肪干细胞复合Ⅰ型胶原海绵多孔支架进行体外培养.将自体脂肪干细胞复合Ⅰ型胶原海绵多孔支架移植到大鼠右上侧乳腺(实验组),将Ⅰ型胶原海绵多孔支架移植到大鼠左上侧乳腺(对照组),共移植6例.12周后观测有无新生脂肪形成;测量新生脂肪组织湿质量;组织切片行HE及油红0染色.结果 SD大鼠脂肪干细胞增殖能力强;具有成骨及成脂分化能力;体外培养2周,胶原支架内可见ASC生长.实验组可见乳腺与胸肌之间有脂肪样新生组织形成,平均湿质量为(121±9) mg.对照组见乳腺与胸肌之间有胶原海绵样组织形成,平均湿质量为(77±6) mg,两组间湿质量存在统计学差异,P<0.05.HE及油红O染色证实,实验组新生组织主要为成熟脂肪组织;而对照组主要为胶原纤维.结论 自体脂肪干细胞复合Ⅰ型胶原海绵多孔支架可在大鼠乳腺区域形成脂肪组织.

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abstractsObjective To explore the feasibility of building the tissue-engineered adipose tissue of breast with adipose- derived stem cells (ASCs) and collagen sponge scaffolds.Methods Enzymatic digestion and centrifugalization were used to isolate ASCs from the inguinal fat pad in adult female Sprague Dawley rats (n=6).After expansion in culture medium,ASCs underwent adipogenic and osteogenic induction and were identified with oil red O staining and yon Kossa staining.For in vitro study,ASCs were seeded onto collagen scaffolds and cultured for 14 days.For in vivo study,ASCs were combined with collagen type Ⅰ sponge scaffolds before implantation.The 6 SD rats underwent implantation of autologous ASCs with collagen sponge into the right side of the upper breast (experimental group),and implantation of collagen sponge into the left side of the upper breast (control group).Twelve weeks later the rats were euthanatized to harvest the implants for wet weight measurement.HE and oil red O staining were used for morphological observation.Results Adipose tissue-like new-born tissues were found in the implantation sites in the experimental group,while collagen sponge-like new-born tissues were found in the implantation sites in the control group.The average wet-weight of the new-born tissue in the experimental group was (121 ±9) mg,significantly heavier than that in the control group [ (77±6) mg,P<0.05].HE staining and oil red O staining confirmed that most of the new-born tissues in the experimental group were mature adipose tissues,while most of the new-born tissues in the control group were collagen fibers.Conclusion Autologous ASCs combined with collagen type Ⅰ scaffolds can be implanted into the breast to build mature adipose tissue in rats.

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