Serban Monica A, Scott Anna, Prestwich Glenn D
Department of Medicinal Chemistry and Center for Therapeutic Biomaterials, The University of Utah, Salt Lake City, Utah, USA.
Curr Protoc Cell Biol. 2008 Sep;Chapter 10:Unit 10.14. doi: 10.1002/0471143030.cb1014s40.
The practice of in vitro three-dimensional (3-D) cell culture has lagged behind the realization that classical two-dimensional (2-D) culture on plastic surfaces fails to mirror normal cell biology. Biologically, a complex network of proteins and proteoglycans that constitute the extracellular matrix (ECM) surrounds every cell. To recapitulate the normal cellular behavior, scaffolds (ECM analogs) that reconstitute the essential biological cues are required. This unit describes the 3-D cell culture and tumor engineering applications of Extracel, a novel semisynthetic ECM (sECM), based on cross-linked derivatives of hyaluronan and gelatin. A simplified cell encapsulation and pseudo-3-D culturing (on top of hydrogels) protocol is provided. In addition, the use of this sECM as a vehicle to obtain tumor xenografts with improved take rates and tumor growth is presented. These engineered tumors can be used to evaluate anticancer therapies under physiologically relevant conditions.
体外三维（3-D）细胞培养的实践落后于人们对以下事实的认识：在塑料表面进行的经典二维（2-D）培养无法反映正常细胞生物学特性。从生物学角度来看，每个细胞周围都有一个由蛋白质和蛋白聚糖构成的复杂网络，即细胞外基质（ECM）。为了重现正常的细胞行为，需要构建能够重构基本生物学信号的支架（ECM类似物）。本单元介绍了Extracel（一种新型半合成ECM，即sECM）在三维细胞培养和肿瘤工程中的应用，它基于透明质酸和明胶的交联衍生物。文中提供了一种简化的细胞封装和伪三维培养（在水凝胶上）方案。此外，还介绍了使用这种sECM作为载体来获得具有更高接种率和肿瘤生长的肿瘤异种移植模型。这些工程化肿瘤可用于在生理相关条件下评估抗癌疗法。
