Davis George E, Kim Dae Joong, Meng Chun-Xia, Norden Pieter R, Speichinger Katherine R, Davis Matthew T, Smith Annie O, Bowers Stephanie L K, Stratman Amber N
Department of Medical Pharmacology and Physiology, University of Missouri School of Medicine and Dalton Cardiovascular Research Center, Columbia, MO, USA.
Methods Mol Biol. 2013;1066:17-28. doi: 10.1007/978-1-62703-604-7_2.
An important advance using in vitro EC tube morphogenesis and maturation models has been the development of systems using serum-free defined media. Using this approach, the growth factors and cytokines which are actually necessary for these events can be determined. The first model developed by our laboratory was such a system where we showed that phorbol ester was needed in order to promote survival and tube morphogenesis in 3D collagen matrices. Recently, we have developed a new system in which the hematopoietic stem cell cytokines, stem cell factor (SCF), interleukin-3 (IL-3), and stromal derived factor-1α (SDF-1α) were added in conjunction with FGF-2 to promote human EC tube morphogenesis in 3D collagen matrices under serum-free defined conditions. This new model using SCF, IL-3, SDF-1α, and FGF-2 also works well following the addition of pericytes where EC tube formation occurs, pericytes are recruited to the tubes, and vascular basement membrane matrix assembly occurs following EC-pericyte interactions. In this chapter, we describe several in vitro assay models that we routinely utilize to investigate the molecular requirements that are critical to EC tube formation and maturation events in 3D extracellular matrix environments.
利用体外内皮细胞（EC）管形态发生和成熟模型取得的一项重要进展是开发了使用无血清限定培养基的系统。采用这种方法，可以确定这些过程实际所需的生长因子和细胞因子。我们实验室开发的第一个模型就是这样一个系统，在该系统中我们发现，为了促进三维胶原基质中的存活和管形态发生，需要佛波酯。最近，我们开发了一种新系统，在无血清限定条件下，将造血干细胞细胞因子、干细胞因子（SCF）、白细胞介素-3（IL-3）和基质衍生因子-1α（SDF-1α）与FGF-2联合添加，以促进三维胶原基质中的人EC管形态发生。在添加周细胞后，这个使用SCF、IL-3、SDF-1α和FGF-2的新模型也能很好地发挥作用，在EC管形成的地方，周细胞被招募到这些管中，并且在EC-周细胞相互作用后发生血管基底膜基质组装。在本章中，我们描述了几种体外分析模型，我们经常利用这些模型来研究在三维细胞外基质环境中对EC管形成和成熟过程至关重要的分子需求。
