Fibrin glue is a candidate scaffold for long-term therapeutic protein expression in spontaneously differentiated adipocytes in vitro
Fibrin glue is a candidate scaffold for long-term therapeutic protein expression in spontaneously differentiated adipocytes in vitro
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DOI：
10.1016/j.yexcr.2011.10.007
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发表时间：
2012-01-01
影响因子：
3.7
通讯作者：
Bujo, Hideaki
中科院分区：
文献类型：
--
作者：
Aoyagi, Yasuyuki;Kuroda, Masayuki;Bujo, Hideaki
脂肪组织有望通过自体移植为蛋白质替代疗法提供细胞来源。然而，移植脂肪细胞周围环境对其长期存活和蛋白质分泌特性的调节尚未确定。我们最近开发了一种前脂肪细胞、天花板培养衍生的增殖脂肪细胞（ccdPA）的制备程序，作为适合稳定基因产物分泌的治疗基因载体。我们在此报告了使用纤维蛋白胶作为移植的 ccdPA 支架在三维培养系统中表达转导基因的评估结果。 ccdPA 分泌从外源转导基因翻译的功能蛋白以及生理脂肪细胞蛋白，ccdPA 的长期生存能力（长达 84 天）取决于纤维蛋白原浓度。 ccdPA 自发积累脂滴，并且其转导的外源基因及其产物的表达水平维持至少 56 天。纤维蛋白原浓度改变ccdPA的脂肪形成分化及其外源基因表达水平，并且不同纤维蛋白原浓度下的外源转导基因表达水平取决于凝胶中脂肪形成分化的程度。这些结果表明，纤维蛋白胶有助于在3D培养系统中传代后维持培养的脂肪细胞的高成脂潜力，并表明一旦它们成功植入移植位点，细胞就会表现出转导基因的表达增加和成脂分化。 (C) 2011 Elsevier Inc. 保留所有权利。
Adipose tissue is expected to provide a source of cells for protein replacement therapies via autotransplantation. However, the conditioning of the environment surrounding the transplanted adipocytes for their long-term survival and protein secretion properties has not been established. We have recently developed a preparation procedure for preadipocytes, ceiling culture-derived proliferative adipocytes (ccdPAs), as a therapeutic gene vehicle suitable for stable gene product secretion. We herein report the results of our evaluation of using fibrin glue as a scaffold for the transplanted ccdPAs for the expression of a transduced gene in a three-dimensional culture system. The ccdPAs secreted the functional protein translated from an exogenously transduced gene, as well as physiological adipocyte proteins, and the long viability of ccdPAs (up to 84 days) was dependent on the fibrinogen concentrations. The ccdPAs spontaneously accumulated lipid droplets, and their expression levels of the transduced exogenous gene with its product were maintained for at least 56 days. The fibrinogen concentration modified the adipogenic differentiation of ccdPAs and their exogenous gene expression levels, and the levels of exogenously transduced gene expression at the different fibrinogen concentrations were dependent on the extent of adipogenic differentiation in the gel. These results indicate that fibrin glue helps to maintain the high adipogenic potential of cultured adipocytes after passaging in a 3D culture system, and suggests that once they are successfully implanted at the transplantation site, the cells exhibit increased expression of the transduced gene with adipogenic differentiation. (C) 2011 Elsevier Inc. All rights reserved.