Syed Sana, Karadaghy Amin, Zustiak Silviya
Biomedical Engineering Department, Saint Louis University.
Biomedical Engineering Department, Saint Louis University;
J Vis Exp. 2015 Mar 25(97):52643. doi: 10.3791/52643.
Currently, most of the in vitro cell research is performed on rigid tissue culture polystyrene (~1 GPa), while most cells in the body are attached to a matrix that is elastic and much softer (0.1-100 kPa). Since such stiffness mismatch greatly affects cell responses, there is a strong interest in developing hydrogel materials that span a wide range of stiffness to serve as cell substrates. Polyacrylamide gels, which are inexpensive and cover the stiffness range of all soft tissues in the body, are the hydrogel of choice for many research groups. However, polyacrylamide gel preparation is lengthy, tedious, and only suitable for small batches. Here, we describe an assay which by utilizing a permanent flexible plastic film as a structural support for the gels, enables the preparation of polyacrylamide gels in a multiwell plate format. The technique is faster, more efficient, and less costly than current methods and permits the preparation of gels of custom sizes not otherwise available. As it doesn't require any specialized equipment, the method could be easily adopted by any research laboratory and would be particularly useful in research focused on understanding stiffness-dependent cell responses.
目前，大多数体外细胞研究是在刚性的组织培养聚苯乙烯（~1吉帕）上进行的，而体内的大多数细胞附着在弹性且柔软得多的基质上（0.1 - 100千帕）。由于这种刚度不匹配会极大地影响细胞反应，因此人们对开发一系列刚度范围的水凝胶材料作为细胞基质有着浓厚的兴趣。聚丙烯酰胺凝胶价格低廉，涵盖了体内所有软组织的刚度范围，是许多研究小组的首选水凝胶。然而，聚丙烯酰胺凝胶的制备过程冗长、繁琐，且仅适用于小批量制备。在此，我们描述了一种测定方法，该方法通过利用永久性柔性塑料薄膜作为凝胶的结构支撑，能够以多孔板形式制备聚丙烯酰胺凝胶。该技术比现有方法更快、更高效且成本更低，并且允许制备其他方法无法获得的定制尺寸的凝胶。由于它不需要任何专门设备，任何研究实验室都可以轻松采用该方法，这对于专注于理解刚度依赖性细胞反应的研究将特别有用。
