Optimizing seeding and culture methods to engineer smooth muscle tissue on biodegradable polymer matrices.
Optimizing seeding and culture methods to engineer smooth muscle tissue on biodegradable polymer matrices.
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DOI：
10.1002/(sici)1097-0290(19980105)57:1
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发表时间：
1998-01
影响因子：
3.8
通讯作者：
Byung‐Soo Kim;A. J. Putnam;T. Kulik;D. Mooney
中科院分区：
文献类型：
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作者：
Byung‐Soo Kim;A. J. Putnam;T. Kulik;D. Mooney
功能性平滑肌（SM）组织的工程化是至关重要的，如果人们希望成功地取代大量的组织含有SM组件与工程等效物。本研究报告了SM细胞（SMC）接种和培养条件对使用聚乙醇酸（PGA）纤维的生物可降解基质（5 × 5 mm，2 mm厚）工程化的SM组织的细胞结构和组成的影响。通过将细胞悬浮液注射到组织培养皿中的聚合物基质中（静态接种），通过在转瓶中搅拌聚合物基质和细胞悬浮液（搅拌接种），或通过用定轨振荡器在管中搅拌聚合物基质和细胞悬浮液（搅拌接种）来接种细胞。粘附于这些基质的SMC的密度是接种溶液中细胞浓度的函数，但在所有条件下，动态接种法与静态接种法相比，粘附于基质的SMC的数量更大（约1个数量级）且分布更均匀。与静态方法相比，动态接种方法也最终导致具有更高细胞构成、更均匀细胞分布和更大弹性蛋白沉积的新组织。接下来通过在搅拌生物反应器中培养细胞-聚合物构建体对比静态培养条件来研究培养条件的影响。5周后，SMC接种的聚合物基质的搅拌培养导致组织的细胞密度为6.4 +/- 0.8 x 10（8）个细胞/cm 3，而静态培养为2.0 +/- 1.1 x 10（8）个细胞/cm 3。弹性蛋白和胶原蛋白的合成速率和沉积在工程组织内也增加了在生物反应器中的培养。在搅拌生物反应器中培养5周后的弹性蛋白含量为24 +/-3%，并且这些组织的弹性蛋白含量和细胞构成与天然SM组织的弹性蛋白含量和细胞构成相当。当动态接种的聚合物基质植入大鼠体内不同时间时，体内也产生了新的组织。总之，由这些研究定义的系统显示出在许多方面与天然SM相当的组织工程化的前景。这种工程组织可能会发现临床应用，并提供一个工具，研究血管发育的分子机制。
The engineering of functional smooth muscle (SM) tissue is critical if one hopes to successfully replace the large number of tissues containing an SM component with engineered equivalents. This study reports on the effects of SM cell (SMC) seeding and culture conditions on the cellularity and composition of SM tissues engineered using biodegradable matrices (5 x 5 mm, 2-mm thick) of polyglycolic acid (PGA) fibers. Cells were seeded by injecting a cell suspension into polymer matrices in tissue culture dishes (static seeding), by stirring polymer matrices and a cell suspension in spinner flasks (stirred seeding), or by agitating polymer matrices and a cell suspension in tubes with an orbital shaker (agitated seeding). The density of SMCs adherent to these matrices was a function of cell concentration in the seeding solution, but under all conditions a larger number (approximately 1 order of magnitude) and more uniform distribution of SMCs adherent to the matrices were obtained with dynamic versus static seeding methods. The dynamic seeding methods, as compared to the static method, also ultimately resulted in new tissues that had a higher cellularity, more uniform cell distribution, and greater elastin deposition. The effects of culture conditions were next studied by culturing cell-polymer constructs in a stirred bioreactor versus static culture conditions. The stirred culture of SMC-seeded polymer matrices resulted in tissues with a cell density of 6.4 +/- 0.8 x 10(8) cells/cm3 after 5 weeks, compared to 2.0 +/- 1.1 x 10(8) cells/cm3 with static culture. The elastin and collagen synthesis rates and deposition within the engineered tissues were also increased by culture in the bioreactors. The elastin content after 5-week culture in the stirred bioreactor was 24 +/- 3%, and both the elastin content and the cellularity of these tissues are comparable to those of native SM tissue. New tissues were also created in vivo when dynamically seeded polymer matrices were implanted in rats for various times. In summary, the system defined by these studies shows promise for engineering a tissue comparable in many respects to native SM. This engineered tissue may find clinical applications and provide a tool to study molecular mechanisms in vascular development.