Bone grafts engineered from human adipose-derived stem cells in perfusion bioreactor culture.
Bone grafts engineered from human adipose-derived stem cells in perfusion bioreactor culture.
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DOI：
10.1089/ten.tea.2009.0164
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发表时间：
2010-01
影响因子：
--
通讯作者：
Vunjak-Novakovic G
中科院分区：
文献类型：
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作者：
Fröhlich M;Grayson WL;Marolt D;Gimble JM;Kregar-Velikonja N;Vunjak-Novakovic G
我们报告了使用人脂肪源性干细胞（hASC）、脱细胞骨支架和灌注生物反应器在体外创建的半厘米大小的骨结构的工程化。hASC容易获得，可以以自体方式使用，在培养中快速扩增，并且能够成骨分化。对来自四个供体的hASC的成骨能力进行表征，并将一个代表性细胞群用于组织工程实验。将培养扩增的hASCs接种在完全脱细胞的天然骨支架（4 mm × 4 mm）上，为成骨分化提供必要的结构和机械环境，并在具有培养基灌注的生物反应器中培养。将间质流速设定为通过增强的质量转运在整个构建体体积（400 μs-1）中维持细胞活力和功能所需的水平。培养5周后，向培养基中添加成骨补充剂（地塞米松、β-甘油磷酸钠、抗坏血酸-2-磷酸盐）显著增加了构建体细胞结构和骨基质组分（胶原、骨唾液酸蛋白、骨骨桥蛋白）的量。培养基灌注显著改善了细胞和骨基质在工程化结构中的分布。总之，hASC、脱细胞骨支架、灌注培养和成骨补充剂的组合导致形成致密且有活力的骨组织构建体。
We report engineering of half centimeter size bone constructs created in vitro using human Adipose-derived Stem Cells (hASC), decellularized bone scaffolds and perfusion bioreactors. The hASCs are easily accessible, can be used in an autologous fashion, are rapidly expanded in culture, and are capable of osteogenic differentiation. hASCs from four donors were characterized for their osteogenic capacity, and one representative cell population was used for tissue engineering experiments. Culture-expanded hASCs were seeded on fully decellularized native bone scaffolds (4 mm Ø × 4 mm) providing the necessary structural and mechanical environment for osteogenic differentiation, and cultured in bioreactors with medium perfusion. The interstitial flow velocity was set to a level necessary to maintain cell viability and function throughout the construct volume (400 μs−1), via enhanced mass transport. After 5 weeks of cultivation, the addition of osteogenic supplements (dexamethasone, sodium-β-glycerophosphate, ascorbic acid-2-phosphate) to culture medium significantly increased the construct cellularity and the amounts of bone matrix components (collagen, bone sialoprotein, bone osteopontin). Medium perfusion markedly improved the distribution of cells and bone matrix in engineered constructs. In summary, a combination of hASCs, decellularized bone scaffold, perfusion culture and osteogenic supplements resulted in the formation of compact and viable bone tissue constructs.