系统背景-因为准确的研究需要组织，而不仅仅是细胞：

尽管传统的单层细胞培养代表了一种成熟且广泛使用的体外方法，但缺乏组织结构和这种模型的复杂性无法为体内的真实生物过程提供信息。世联博研北京公司提供的美国flexcell tissue train 3D动态机械力微环境三维培养构建系统的细胞培养技术的新进展通过创建强大的三维 （3D） 模型来概括原代组织的细胞异质性、结构和功能，彻底改变了生物医学研究的体外培养工具。这些模型还弥合了传统二维 （2D） 单层培养物与动物模型之间的差距。3D 培养系统使研究人员能够在一个培养皿中重建人体器官和疾病，因此在再生医学、药物发现、精准医学和癌症研究以及基因表达研究等许多应用中具有很大的前景。

美国flexcell tissue train 3D动态机械力微环境三维培养构建系统技术使您能够在几分钟内创建高度受控的 3D 组织培养模型。 可重现、成本效益高且对人类生物学具有高度预测性。

系统介绍：

可在基底硬度刚度模量0.1-80kpa范围内对二维、三维细胞和组织各种培养物提供轴向和圆周应力加载。

基底微图案纳米表面，该图案具有亚微米对齐的凹槽，并且其硬度可调节（0.1-80kpa）。 微图案有助于形成排列的细胞单层，使它们具有结构排列和组织性，从而使细胞更加成熟和生理地表达。

产品成熟：文献达4000多篇，国内有150多家成功单位使用。

亮点：

1）该系统对二维、三维细胞和组织各种培养物提供轴向和圆周应力加载；不但具有双轴向拉伸力加载，还具备单轴向加力功能

2）计算机控制的应力加载系统，为体外培育的细胞提供j确的、可控制的、可重复的、静态的或者周期性的应力变化。

3）使用真空泵，抻拉培养板底部的弹性硅胶模，细胞培养板底部高伸展度可达到33%，通过气体装置可以自动调节和控制应力。

4）基于柔性膜基底变形、受力均匀;

5）可实时观察细胞、组织在应力作用下的反应;

6）具的flexstop隔离阀可使同一块培养板力的一部分培养孔的细胞受力，一部分培养孔的细胞不受力，方便对比实验;

7）与压力传导仪整合，同时兼备多通道细胞压力加载功能;

8）与Flex Flow平行板流室配套，可在牵拉细胞的同时施加流体切应力;

9）多达4通道，可4个不同程序同时运行，进行多个不同拉伸形变率对比实验；

10）同一程序中可以运行多种频率，多种振幅和多种波形；

11）加载模拟波形种类丰富：静态波形、正旋波形、心动波形、三角波形、矩形以及各种te制波形；

12）更好地控制在超低或超高应力下的波形；

13)电脑系统对牵张拉伸力加载周期、大小、频率、持续时间j确智能调控

14)加载分析各种细胞在牵张拉应力刺激下的生物化学反应

15)伸展度范围广：0-33%

16)牵拉频率范围广：0.01-5Hz

三维种子细胞构建人工生物组织系统(Creating a Bioartifical Construct with the Tissue Train System)

点击添加图片描述（多60个字）编辑

特点：

1）对生长在三维状态下的细胞进行静态的或者周期性的牵张拉伸刺激培养，可以进行实时观察分析。

2）对生长在三维环境下的细胞进行单轴向或者双轴向的静态或者周期性的应力加载实验

3)可建立te制的各种模拟实验：心率模拟实验，步行模拟实验，跑动模拟实验和其他动力模拟实验。

4)可构建长度达35mm的生物人工组织

5）具有丰富的三维培养模具和多种蛋白包被材料的自动细胞组织三维培养系统

点击添加图片描述（多60个字）编辑

6）该系统以立体水凝胶为三维培养支架， 水凝胶支架具有大量体内微环境基质的特征，水凝胶所具有的三维网络结构、含水量高和力学性能可控等特性与体内细胞所处基质微环境相似, 被广泛用于工程化组织的体外构建研究，水凝胶的硬度调控范围很大, 有利于模拟体内生理或病理力学微环境

是真正意义上的三维培养

适用范围

1）FLEXCELL的Tissue Train ?培养体系，是为了解决这一组织培养过程中的难题，这个培养体系通过为细胞和基质提供三维支架矩阵组织、动态的拉伸力和多种几何模型来创建不同形状的生物人工组织（如线性，梯形和圆形）。

2水凝胶基质力学环境模拟

3）生物材料的细胞力学微环境体外构建系统

4）基于干细胞3D力学环境的工程化微组织构建研究

Tissue Train ?培养系统应用背景

体外培养在与真实组织在结构上和功能上相似的人工组织需要以下几个基本条件：

（1）细胞

（2）支架矩阵组织

（3）培养基和生长因子和（4）机械刺激。这些条件彼此相互影响，并且相互之间共同来促进形成能够承受生物机械力的，且结构比较稳定的组织。而在人工组成形成的过程中，这些细胞按照发育途径形成具有一定几何形状的细胞外基质结构。其中一些信号转导途径参与了细胞外基质组合物的形成。这些途径中，有些是由细胞基质的机械变形调节，并通过膜结合蛋白，如整合素，粘着斑复合体，细胞粘附分子和离子通道传递到细胞内。这些途径中细胞还可以响应配体，如细胞基质形变所释放的细胞因子，激素或生长因子等。

为了维持肌肉骨骼组织的完整性和强度，组织内细胞需要保持一定水平的的内在应力。如果缺乏这种内在的应力，组织会缺少强度导致细胞结构的破坏或者组织的断裂。目前一般认为如果在固定四肢，卧床休息或在内在应力水平的降低的情况下，将导致骨中矿物质流失，骨组织萎缩，骨骼弱化，以及合成代谢活性的降低和分解代谢活性的增加。

为了在体外培养与原生组织类似的人工组织，重要的就是能够创建模拟体内条件的环境。细胞在具有机械运动作用的的环境中培养，可以促进细胞的新陈代谢，并可以改变细胞的形状和其它性能。因此，在体外形成过程中建立和保持一个具备机械作用的环境（即张力，剪切力或压缩）就成为这一过程中至关重要的。除了具备机械作用的环境，在三维环境下培养细胞可以比静态二维培养法更好地模拟原生环境。

典型应用文献摘选：

1. Abraham T, Kayra D, McManus B, Scott A. Quantitative assessment of forward and backward second harmonic three dimensional images of collagen type I matrix remodeling in a stimulated cellular environment. J Struct Biol 180(1):17-25, 2012.

2. Ahearne M, Bagnaninchi PO, Yang Y, El Haj AJ. Online monitoring of collagen fibre alignment in tissue-engineered tendon by PSOCT. J Tissue Eng Regen Med 2(8):521-524, 2008.

3. Allison DA, Wight TN, Ripp NJ, Braun KR, Grande-Allen KJ. Endogenous overexpression of hyaluronan synthases within dynamically cultured collagen gels: implications for vascular and valvular disease. Biomaterials 29:2969-2976, 2008.

4. Barbolina MV, Liu Y, Gurler H, Kim M, Kajdacsy-Balla AA, Rooper L, Shepard J, Weiss M, Shea LD, Penzes P, Ravosa MJ, Stack MS. Matrix rigidity activates Wnt signaling through down-regulation of Dickkopf-1 protein. J Biol Chem 288(1):141-51, 2013.

5. Bertrand AT, Ziaei S, Ehret C, Duchemin H, Mamchaoui K, Bigot A, Mayer M, Quijano-Roy S, Desguerre I, Lainé J, Ben Yaou R, Bonne G, Coirault C. Cellular microenvironments reveal defective mechanosensing responses and elevated YAP signaling in LMNA-mutated muscle precursors. J Cell Sci 127(Pt 13):2873-84, 2014.

6. Cao TV, Hicks MR, Campbell D, Standley PR. Dosed myofascial release in three-dimensional bioengineered tendons: effects on human fibroblast hyperplasia, hypertrophy, and cytokine secretion. J Manipulative Physiol Ther 36(8):513-21, 2013.

7. Cao TV, Hicks MR, Zein-Hammoud M, Standley PR. Duration and magnitude of myofascial release in 3-dimensional bioengineered tendons: effects on wound healing. J Am Osteopath Assoc 115(2):72-82, 2015.

8. Charoenpanich A, Wall ME, Tucker CJ, Andrews DM, Lalush DS, Loboa EG. Microarray analysis of human adipose-derived stem cells in three-dimensional collagen culture: osteogenesis inhibits bone morphogenic protein and Wnt signaling pathways, and cyclic tensile strain causes upregulation of proinflammatory cytokine regulators and angiogenic factors. Tissue Eng Part A 17(21-22):2615-2627, 2011.

9. Clause KC, Tinney JP, Liu LJ, Gharaibeh B, Huard J, Kirk JA, Shroff SG, Fujimoto KL, Wagner WR, Ralphe JC, Keller BB, Tobita K. A three-dimensional gel bioreactor for assessment of cardiomyocyte induction in skeletal muscle-derived stem cells. Tissue Eng Part C Methods 16(3):375-385, 2010.

10. Clause KC, Tinney JP, Liu LJ, Keller BB, Tobita K. Engineered early embryonic cardiac tissue increases cardiomyocyte proliferation by cyclic mechanical stretch via p38-MAP kinase phosphorylation. Tissue Engineering Part A 15(6):1373-1380, 2009.

11. Clause KC, Tinney JP, Liu JL, Keller BB, Huard J, Tobita K. p38MAP-kinase regulates cardiomyocyte proliferation and contractile properties of engineered early embryonic cardiac tissue [abstract]. Weinstein Cardiovascular Development Research Conference, Indianapolis, IN, 2007.

12. Clause KC, Tinney JP, Liu JL, Gharaibeh B, Fujimoto LK, Wagner WR, Ralphe JC, Keller BB, Huard J, Tobita K. Functioning engineered cardiac tissue from skeletal muscle derived stem cells [abstract]. 4th Annual Symposium of AHA Council on Basic Cardiovascular Sciences, Keystone CO, 2007.

13. de Jonge N, Foolen J, Brugmans MC, S?ntjens SH, Baaijens FP, Bouten CV. Degree of scaffold degradation influences collagen (re)orientation in engineered tissues. Tissue Eng Part A 20(11-12):1747-57, 2014.

14. de Lange WJ, Grimes AC, Hegge LF, Ralphe JC. Ablation of cardiac myosin-binding protein-C accelerates contractile kinetics in engineered cardiac tissue. J Gen Physiol 141(1):73-84, 2013.

15. Ferdous Z, Lazaro LD, Iozzo RV, H??k M, Grande-Allen KJ. Influence of cyclic strain and decorin deficiency on 3D cellularized collagen matrices. Biomaterials 29(18):2740-2748, 2008.

16. Freeman SA, Christian S, Austin P, Iu I, Graves ML, Huang L, Tang S, Coombs D, Gold MR, Roskelley CD. Applied stretch initiates directional invasion through the action of Rap1 GTPase as a tension sensor. J Cell Sci 130(1):152-163, 2017.

17. Garvin J, Qi J, Maloney M, Banes AJ. Novel system for engineering bioartificial tendons and application of mechanical load. Tissue Eng 9(5):967-979, 2003.

18. Henshaw DR, Attia E, Bhargava M, Hannafin JA. Canine ACL fibroblast integrin expression and cell alignment in response to cyclic tensile strain in three-dimensional collagen gels. J Orthop Res 24(3):481-490, 2006.

19. Huang G, Wang L, Wang S, Han Y, Wu J, Zhang Q, Xu F, Lu TJ. Engineering three-dimensional cell mechanical microenvironment with hydrogels. Biofabrication 4(4):042001, 2012.

20. Jobling AI, Gentle A, Metlapally R, McGowan BJ, McBrien NA. Regulation of scleral cell contraction by transforming growth factor-? and stress: competing roles in myopic eye growth. J Biol Chem 284(4):2072-2079, 2009.

21. Jones ER, Jones GC, Legerlotz K, Riley GP. Cyclical strain modulates metalloprotease and matrix gene expression in human tenocytes via activation of TGFβ. Biochim Biophys Acta 1833(12):2596-2607, 2013.

22. Lee CH, Shin HJ, Cho IH, Kang YM, Kim IA, Park KD, Shin JW. Nanofiber alignment and direction of mechanical strain affect the ECM production of human ACL fibroblast. Biomaterials 26(11):1261-1270, 2005.

23. Masumoto H, Nakane T, Tinney JP, Yuan F, Ye F, Kowalski WJ, Minakata K, Sakata R, Yamashita JK, Keller BB. The myocardial regenerative potential of three-dimensional engineered cardiac tissues composed of multiple human iPS cell-derived cardiovascular cell lineages. Sci Rep 6:29933, 2016.

24. Nguyen MD, Tinney JP, Ye F, Elnakib AA, Yuan F, El-Baz A, Sethu P, Keller BB, Giridharan GA. Effects of physiologic mechanical stimulation on embryonic chick cardiomyocytes using a microfluidic cardiac cell culture model. Anal Chem 87(4):2107-13, 2015.

25. Nieponice A, Maul TM, Cumer JM, Soletti L, Vorp DA. Mechanical stimulation induces morphological and phenotypic changes in bone marrow-derived progenitor cells within a three-dimensional fibrin matrix. J Biomed Mater Res A 81(3):523-530, 2007.

26. Nourse MB, Halpin DE, Scatena M, Mortisen DJ, Tulloch NL, Hauch KD, Torok-Storb B, Ratner BD, Pabon L, Murry CE. VEGF induces differentiation of functional endothelium from human embryonic stem cells: implications for tissue engineering. Arterioscler Thromb Vasc Biol 30(1):80-89, 2010.

27. Peters AS, Brunner G, Krieg T, Eckes B. Cyclic mechanical strain induces TGFβ1-signalling in dermal fibroblasts embedded in a 3D collagen lattice. Arch Dermatol Res 307(2):191-7, 2015.

28. Qi J, Chi L, Bynum D, Banes AJ. Gap junctions in IL-1β-mediated cell survival response to strain. J Appl Physiol 110(5):1425-1431, 2011.

29. Qi J, Chi L, Faber J, Koller B, Banes AJ. ATP reduces gel compaction in osteoblast-populated collagen gels. J Appl Physiol 102(3):1152-60, 2007.

30. Qi J, Chi L, Maloney M, Yang X, Bynum D, Banes AJ. Interleukin-1? increases elasticity of human bioartificial tendons. Tissue Eng 12(10):2913-2925, 2006.

31. Qi J, Fox AM, Alexopoulos LG, Chi L, Bynum D, Guilak F, Banes AJ. IL-1??decreases the elastic modulus of human tenocytes. J Appl Physiol 101(1):189-95, 2006.

32. Qi J, Chi L, Wang J, Sumanasinghe R, Wall M, Tsuzaki M, Banes AJ. Modulation of collagen gel compaction by extracellular ATP is MAPK and NF-?B pathways dependent. Exp Cell Res 315(11):1990-2000, 2009.

33. Rathbone SR, Glossop JR, Gough JE, Cartmell SH. Cyclic tensile strain upon human mesenchymal stem cells in 2D and 3D culture differentially influences CCNL2, WDR61 and BAHCC1 gene expression levels. J Mech Behav Biomed Mater 11:82-91, 2012.

34. Raval KK, Tao R, White BE, De Lange WJ, Koonce CH, Yu J, Kishnani PS, Thomson JA, Mosher DF, Ralphe JC, Kamp TJ. Pompe disease results in a Golgi-based glycosylation deficit in human induced pluripotent stem cell-derived cardiomyocytes. J Biol Chem 290(5):3121-36, 2015.

35. Ruan JL, Tulloch NL, Saiget M, Paige SL, Razumova MV, Regnier M, Tung KC, Keller G, Pabon L, Reinecke H, Murry CE. Mechanical stress promotes maturation of human myocardium from pluripotent stem cell-derived progenitors. Stem Cells 33(7):2148-57, 2015.

36. Schmidt JB, Chen K, Tranquillo RT. Effects of intermittent and incremental cyclic stretch on ERK signaling and collagen production in engineered tissue. Cellular and Molecular Bioengineering 1-10, 2015.

37. Sumanasinghe RD, Bernacki SH, Loboa EG. Osteogenic differentiation of human mesenchymal stem cells in collagen matrices: effect of uniaxial cyclic tensile strain on bone morphogenetic protein (BMP-2) mRNA expression. Tissue Eng 12(12):3459-3465, 2006.

38. Taylor SE, Vaughan-Thomas A, Clements DN, Pinchbeck G, Macrory LC, Smith RK, Clegg PD. Gene expression markers of tendon fibroblasts in normal and diseased tissue compared to monolayer and three dimensional culture systems. BMC Musculoskelet Disord 10:27, 2009.

39. Tchao J, Han L, Lin B, Yang L, Tobita K. Combined biophysical and soluble factor modulation induces cardiomyocyte differentiation from human muscle derived stem cells. Sci Rep 4:6614, 2014.

40. Tchao J, Kim JJ, Lin B, Salama G, Lo CW, Yang L, Tobita K. Engineered human muscle tissue from skeletal muscle derived stem cells and induced pluripotent stem cell derived cardiac cells. Int J Tissue Eng. 2013:198762, 2013.

41. Tobita K, Liu LJ, Janczewski AM, Tinney JP, Nonemaker JM, Augustine S, Stolz DB, Shroff SG, Keller BB. Engineered early embryonic cardiac tissue retains proliferative and contractile properties of developing embryonic myocardium. Am J Physiol Heart Circ Physiol 291(4):H1829-37, 2006.

42. Tondon A, Haase C, Kaunas R. Mechanical stretch assays in cell culture systems. In: Handbook of Imaging in Biological Mechanics, ed. Neu CP, Genin GM. CRC Press: Boca Raton, 2015.

43. Triantafillopoulos IK, Banes AJ, Bowman KF Jr, Maloney M, Garrett WE Jr, Karas SG. Nandrolone decanoate and load increase remodeling and strength in human supraspinatus bioartificial tendons. Am J Sports Med 32(4):934-943, 2004.

44. Tulloch NL, Muskheli V, Razumova MV, Korte FS, Regnier M, Hauch KD, Pabon L, Reinecke H, Murry CE. Growth of engineered human myocardium with mechanical loading and vascular coculture. Circ Res 109(1):47-59, 2011.

45. Weinbaum JS, Schmidt JB, Tranquillo RT. Combating adaptation to cyclic stretching by prolonging activation of extracellular signal-regulated kinase. Cellular and Molecular Bioengineering 6(3):279-286, 2013.

46. Wen W, Chau E, Jackson-Boeters L, Elliott C, Daley TD, Hamilton DW. TGF-?1 and FAK regulate periostin expression in PDL fibroblasts. J Dent Res 89(12):1439-1443, 2010.

47. Yang G, Rothrauff BB, Lin H, Gottardi R, Alexander PG, Tuan RS. Enhancement of tenogenic differentiation of human adipose stem cells by tendon-derived extracellular matrix. Biomaterials 34(37):9295-306, 2013.

48. Yang Y, Wimpenny I, Wang RK. Application of polarization-sensitive OCT and Doppler OCT in tissue engineering. In: Optical Techniques in Regnerative Medicine, edited by Morgan SP, Rose F, Matcher SJ. Taylor & Francis Group: Florida, p. 307-327, 2014.

49. Ye F, Yuan F, Li X, Cooper N, Tinney JP, Keller BB. Gene expression profiles in engineered cardiac tissues respond to mechanical loading and inhibition of tyrosine kinases. Physiol Rep 1(5):e00078, 2013.