Human lipogenesis with basic fibroblast growth factor gene transfected preadipocytes for breast reconstruction.

人类脂肪生成与碱性成纤维细胞生长因子基因转染前脂肪细胞用于乳房重建。

• 批准号: 17591330
• 负责人: INAMOTO Takashi
• 金额: $ 2.3万
• 依托单位: The Tazuke Kofukai (2006)Kyoto University (2005)
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2005
• 资助国家: 日本
• 起止时间: 2005 至 2006
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-17591330/
• 关键词: fatty tissue; progenitor cell; bFGF; tissue regeneration; breast reconstruction; 遺伝子導入; アデノウイルス

1) Human stromal cells including preadipocytes which were separated from fatty tissue obtained at breast cancer operation were incubated and passed in vitro. Proliferation of stromal cells was preserved through 10^<th> passage. Basic fibroblast growth factor (bFGF) enhanced the proliferation of stromal cells. Cell yield exceeded 1000 times of the cell number at the beginning of culture. Cultured stromal cells maintained maturation activity to adipocyte at the initial level through 3^<rd> passage and declined thereafter. This result suggested that the optimal cell source for lipogenesis was 3^<rd> passage of cultured stromal cells.2) Proliferation of stromal cells was enhanced when they were cutltured with autorogous serum compared with fetal calf serum (FCS). Effusion of axillary cavity after dissection of lymph nodes showed similar effect on the proliferation of stromal cells in vitro as FCS. This result suggested cultured stromal cells would proliferate in vivo.3) 3^<rd> passage of s … More tromal cells cultured with bFGF was implanted with collagen disk scaffold incorporated by the microspheres containing bFGF into subcutaneous tissue of the back of nude mice. Adipogenesis at the implanted site of scaffold was evaluated histologically. Area of angiogenesis in 2 x 10^6 cultured stromal cell implant group was largest among groups implanted different number of cultured stromal cells, but area of adipogenesis expressing human vimentin was less than 1% of whole adipogenesis. In 8 x 10^6 cultured stromal cell implant group, area of human vimentin positive adipose tissue was 15% of whole adiopogenesis, although the area of whole adipogenesis was less than 2 x 10^6 cultured stromal cell implant group.4) Initialially, b-FGF gene transfected using adenovirus vector, and obtained the result that stromal cells transfected b-FGF gene proliferated in vitro without exogenous b-FGF and differentiated into mature adipocytes. For future clinical application of this adipogenesis method, we are investigating a safe and efficient non-viral gene delivery system. Less

1)从乳腺癌手术中获得的脂肪组织中分离出包括前脂肪细胞在内的人基质细胞，体外培养并传代。基质细胞的增殖保持至10<th>代。碱性成纤维细胞生长因子（bFGF）促进基质细胞的增殖。细胞产量超过培养初期细胞数的1000倍。培养的基质细胞在第3代时仍保持着向脂肪细胞成熟的活性<rd>，以后逐渐下降。结果提示，体外培养的基质细胞是脂肪细胞合成的最佳细胞来源。<rd>2）与胎牛血清（FCS）相比，自体血清培养的基质细胞增殖能力更强。淋巴结清扫后的腋窝积液对基质细胞的体外增殖作用与FCS相似。结果表明，培养的基质细胞在体内可以增殖<rd>。 ...更多信息 将bFGF微球包埋的胶原圆盘支架植入裸鼠背部皮下组织，观察bFGF对裸鼠成纤维细胞生长的影响。组织学评价支架植入部位的脂肪生成。在植入不同数量培养基质细胞的组中，2 × 10^6培养基质细胞植入组的血管生成面积最大，但表达人波形蛋白的脂肪生成面积不到整个脂肪生成的1%。在8 × 10^6培养的基质细胞植入组中，人波形蛋白阳性脂肪组织的面积为整个脂肪生成的15%，尽管整个脂肪生成的面积小于2 × 10^6培养的基质细胞植入组。获得了转染b-FGF基因的基质细胞在无外源性b-FGF的条件下体外增殖并分化为成熟脂肪细胞的结果。为了将来这种脂肪生成方法的临床应用，我们正在研究一种安全有效的非病毒基因递送系统。少