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具有机械调节和无损细胞分离功能的细胞培养平台。

Cell culture platform with mechanical conditioning and nondamaging cellular detachment.

机构信息

Department of Biomedical Engineering, Case Western Reserve University, Wickenden 220, 10900 Euclid Avenue,Cleveland, Ohio 44106, USA.

出版信息

J Biomed Mater Res A. 2010 May;93(2):411-8. doi: 10.1002/jbm.a.32754.

Abstract

Cells implanted after injury may remodel undesirably with improper mechanical stimulation from surrounding tissue. Proper conditioning of tissue engineered constructs before implantation can lead to suitable tissue architectures, along with an extracellular matrix (ECM) environment that more closely mimics native tissue. Additionally, cell implantation without bulky polymeric scaffolding is often desirable. Previous researchers have created devices capable of applying mechanical forces to cells (e.g., stretch) but cellular removal from these devices, such as by trypsin, often results in irreversible damage. Conversely, devices are available that can detach intact cells, but these are inelastic, nonstretchable substrates. We have created a cell culture platform that allows for mechanical conditioning and then subsequent nondamaging detachment of those cells. We have modified silicone culture surfaces, to incorporate thermally responsive polymers of N-isopropylacrylamide (NIPAAm) to create an elastic substrate that can also change surface properties with temperature change. A copolymer of NIPAAm and 10percent w/w acrylic acid (AAc) was conjugated to an amine-bonded silicone surface through carbodiimide chemistry. Cells were able to attach to the resulting surfaces at 37 degreeC and showed detachment by rounded morphology at 25degreeC. Following mechanical stretching, cells were still able to spontaneously detach from these modified silicone surfaces with temperature change.

摘要

细胞在损伤后可能会因周围组织的不当机械刺激而进行不理想的重塑。在植入前对组织工程构建体进行适当的调理,可以形成合适的组织结构,并形成更接近天然组织的细胞外基质 (ECM) 环境。此外,通常希望在没有大块聚合物支架的情况下进行细胞植入。以前的研究人员已经开发出能够对细胞施加机械力(例如拉伸)的设备,但从这些设备中取出细胞(例如用胰蛋白酶)通常会导致不可逆的损伤。相反,也有能够分离完整细胞的设备,但这些设备是无弹性的、不可拉伸的基底。我们创建了一个细胞培养平台,允许进行机械调理,然后无损地分离这些细胞。我们已经对硅酮培养表面进行了修改,将 N-异丙基丙烯酰胺 (NIPAAm) 的热响应聚合物结合到弹性基底中,该基底还可以随温度变化改变表面特性。NIPAAm 和 10% w/w 丙烯酸 (AAc) 的共聚物通过碳二亚胺化学与胺键合的硅酮表面结合。细胞在 37°C 时能够附着在这些表面上,并在 25°C 时通过圆形形态显示出分离。在机械拉伸后,细胞仍然能够通过温度变化从这些改性的硅酮表面自发地分离。

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