Department of Bioengineering, University of California San Diego, La Jolla, California, United States of America.
PLoS One. 2010 Sep 27;5(9):e13039. doi: 10.1371/journal.pone.0013039.
The native extracellular matrix (ECM) consists of a highly complex, tissue-specific network of proteins and polysaccharides, which help regulate many cellular functions. Despite the complex nature of the ECM, in vitro cell-based studies traditionally assess cell behavior on single ECM component substrates, which do not adequately mimic the in vivo extracellular milieu.
METHODOLOGY/PRINCIPAL FINDINGS: We present a simple approach for developing naturally derived ECM coatings for cell culture that provide important tissue-specific cues unlike traditional cell culture coatings, thereby enabling the maturation of committed C2C12 skeletal myoblast progenitors and human embryonic stem cells differentiated into cardiomyocytes. Here we show that natural muscle-specific coatings can (i) be derived from decellularized, solubilized adult porcine muscle, (ii) contain a complex mixture of ECM components including polysaccharides, (iii) adsorb onto tissue culture plastic and (iv) promote cell maturation of committed muscle progenitor and stem cells.
This versatile method can create tissue-specific ECM coatings, which offer a promising platform for cell culture to more closely mimic the mature in vivo ECM microenvironment.
天然细胞外基质(ECM)由高度复杂的、组织特异性的蛋白质和多糖网络组成,它有助于调节许多细胞功能。尽管 ECM 的性质复杂,但传统的基于细胞的体外研究通常评估细胞在单个 ECM 成分底物上的行为,而这些底物不能充分模拟体内细胞外环境。
方法/主要发现:我们提出了一种简单的方法来开发用于细胞培养的天然衍生的 ECM 涂层,这些涂层提供了不同于传统细胞培养涂层的重要组织特异性线索,从而使定向的 C2C12 骨骼肌成肌细胞祖细胞和分化为心肌细胞的人胚胎干细胞成熟。在这里,我们表明天然肌肉特异性涂层可以(i)从去细胞化的、可溶解的成年猪肌肉中获得,(ii)包含包括多糖在内的复杂 ECM 成分混合物,(iii)吸附到组织培养塑料上,以及(iv)促进定向肌肉祖细胞和干细胞的细胞成熟。
这种多功能的方法可以创建组织特异性的 ECM 涂层,为更接近模拟成熟体内 ECM 微环境的细胞培养提供了一个有前途的平台。