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組織工程有望實現革命性*的治療,不過,盡管如此,它仍面臨著一些有關的細胞在體外培養(yǎng)的關鍵問題。具體來說,獲得在實驗室培養(yǎng)條件而準確地模仿在體內的生長條件是要解決的關鍵挑戰(zhàn)之一。
在這些情況下,再現一個“人造”血管系統(tǒng)能夠提供充足的氧氣和營養(yǎng)給細胞生長,以及帶走細胞的新陳代謝產生的廢物,是zui困難的問題之一。
在三維支架上進行細胞的培養(yǎng)還是我們要去面對的。假設在開始培養(yǎng)時氧氣和營養(yǎng)物質均勻的分布在整個支架,細胞以相同的速度在整個支架上生長。然而,這些位于支架中心的細胞需要氧氣和營養(yǎng)物質,二氧化碳和廢物發(fā)熱帶走,因為相比細胞的新陳代謝,被動的交換更新的速度太慢。然后,它被發(fā)現通常會導致大于1毫米支架由于缺乏血管供應的可發(fā)育細胞和母體壞死核心的外殼。
因此,改善營養(yǎng)運輸交換在細胞群體三維體外實驗中至關重要的,為整個支架(灌注)培養(yǎng)介質施加一定的流速已經實驗證明非常有效的比靜態(tài)培養(yǎng)。
同樣,流體刺激對某些種類的組織,例如,血管組織適當的發(fā)展也至關重要。在這種情況下,內膜層內皮細胞??的強烈影響血流量的作用,在一般情況下,整個管路的流動已被實驗證明施加的強烈影響這些細胞的行為。因此,血管流量條件下的培養(yǎng)是至關重要的,因為沒有其他條件,更好地重現細胞在體內所經歷的條件。
軟骨
Lin et al. Chondrocytes culture in three-dimensional porous alginate scaffolds enhanced cell proliferation, matrix synthesis and gene expression. J Biomed Mater Res A 88 (2009) 23-33.
Raimondi et al. Engineered cartilage constructs subject to very low regimes of interstitial perfusion. Biorheology 45 (2008) 471-478
Wendt et al. Uniform tissues engineered by seeding and culturing cells in 3D scaffolds under perfusion at defined oxygen tensions. Biorrheology 43 (2006) 481-488
Mahmoudifar et al. Effect of seeding and bioreactor culture conditions on the development of human tissue-engineered cartilage. Tissue Engineering 12 (2006) 1675-1685
骨
Grayson et al. Engineering anatomically shaped human bone grafts. PNAS (2009)
Fröhlich et al. Bone grafts engineered from human adipose-derived stem cells in perfusion bioreactor culture. Tissue Eng A (2009)
Du et al. Oscillatory perfusion of CaP-based tissue engineered bone with and without dexamethasone. Annals of Biomedical Engineering 37 (2009) 146-155
Bernhardt et al. Proliferation and osteogenic differentiation of human bone marrow stromal cells on alginate-gelatine-hydroxyapatite scaffolds with anisotropic pore structure. Journal of Tissue Engineering and Regenerative Medicine 3 (2009) 54-62
血管
Bjork et al. Transmural flow bioreactor for vascular tissue engineering. Biotechnol Bioeng 15 (2009) 1197-1206
McIlhenny et al. Linear shear conditioning improves vascular graft retention of adipose-derived stem cells by up-regulation of the α5β1 integrin. Tissue Eng A (2009)
Mertsching et al. Bioreactor technology in cardiovascular tissue engineering. Adv Biochem Eng Biotechnol (2008)
Mironov et al. Cardiovascular tissue engineering I. Perfusion bioreactors: a review. J Long Term Eff Med Implants 16 (2006) 111-130.
*
Ruel et al. A new bioreactor for the development of tissue-engineered heart valves. Annals of Biomedical Engineering 37 (2009) 674-681.
Ott et al. Perfusion-decellularized matrix: using nature’s platform to engineer a bioartificial heart. Nature Medicine 14 (2008) 213-221
Brown et al. Pulsatile perfusion bioreactor for cardiac tissue engineering. Biotechnol Prog 24 (2008) 907-920
Gulbins et al. A low-flow adaptation phase improves shear-stress resistance of artificially seeded endothelial cells. Thorac Cardiovasc Surg 53 (2005) 96-102
肝臟
Atala et al. Engineering complex tissues. Sci Transl Med 14 (2012)
Li et al. Cells and Materials for Liver Tissue Engineering. Cell Transplant (2012)
Yagi et al. Decellularized scaffold as a platform for novel regenerative therapy. Nihon Geka Gakkai Zasshi 113 (2012) 419-423
Uygun et al. Application of whole-organ tissue engineering in hepatology. Nat Rev Gastroenterol Hepatol(2012
膀胱
Yagi et al. Whole-organ re-engineering: a regenerative medicine approach in digestive surgery for organ replacement. Surg Today (2012)
Horst et al. Engineering functional bladder tissues. J Tissue Eng Regen Med (2012)
Chen et al. Tissue engineering of bladder using vascular endothelial growth factor gene-modified endothelial progenitor cells. Int J Artif Organs 34 (2011) 1137-1146
Davis et al. Construction and evaluation of urinary bladder bioreactor for urologic tissue-engineering purposes. Urology 78 (2011) 954-960.
MECHANOTRANSDUCTION 力學傳導
Cicha et al. Shear stress preconditioning modulates endothelial susceptibility to circulating TNF-α and monocytic cell recruitment in a simplified model of arterial bifurcations. Atherosclerosis 207 (2009) 93-102
Kowalsky et al. oxLDL facilitates flow-induced realignment of human aortic endothelial cells. Am J Physiol Cell Physiol 295 (2008) C332-340
Cicha et al. Pharmacological inhibition of RhoA signaling prevents connective tissue growth factor induction in endothelial cells exposed to non-uniform shear stress. Atherosclerosis 196 (2008) 136-145
Friedrich et al. Podocytes are sensitive to fluid shear stress in vitro. Am J Physiol Renal Physiol 291 (2006) F856-865
細胞粘附
Fuchs et al. Flow-based measurements of von Willebrand factor (VWF) function: binding to collagen and plaet adhesion under physiological shear rate. Thrombosis Research (2009)
Pfaff et al. Involvement of endothelial ephrin-B2 in adhesion and transmigration of EphB-receptor-expressing monocytes. J Cell Sci 121 (2008) 3842-3850
Petoumenos et al. High density lipoprotein exerts vasculoprotection via endothelial progenitor cells. Journal of Cellular and Molecular Medicine (2008)
MICROFLUIDICS 微灌注
Huang et al. An integrated microfluidic cell culture system for high-throughput perfusion three-dimensional cell culture-based assays: effect of cell culture model on the results of chemosensitivity assays. Lab Chip 13 (2013):1133-43
Kim et al. Engineering of functional, perfusable 3D microvascular networks on a chip facilitates flow-induced realignment of human aortic endothelial cells. Lab Chip 13 (2013): 1489-1500
Xu et al. Application of a microfluidic chip-based 3D co-culture to test drug sensitivity for individualized treatment of lung cancer. Biomaterials 34 (2013):4109-17
Yokokawa et al. A perfusable microfluidic device with on-chip total internal reflection fluorescence microscopy (TIRFM) for in situ and real-time monitoring of live cells. Biomed Microdevices 14 (2012):791-7
細胞動態(tài)種植
Álvarez-Barreto et al. Flow Perfusion Improves Seeding of Tissue Engineering Scaffolds with Different Architectures. Annals of Biomedical Engineering, 35 (2007): 429-442
Maidhof et al. Perfusion Seeding of Channeled Elastomeric Scaffolds with Myocytes and Endothelial Cells for Cardiac Tissue Engineering. Biotechnol. Prog. 26 (2010): 565-572
Kock et al. Perfusion cell seeding on large porous PLA/calcium phosphate composite scaffolds in a perfusion bioreactor system under varying perfusion parameters. J Biomed Mater Res Part A: 95A (2010): 1011–1018.
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