Kroeze Robert Jan, Knippenberg Marlene, Helder Marco N
Department of Orthopaedic Surgery, VU University Medical Center, Amsterdam, The Netherlands.
Methods Mol Biol. 2011;702:233-48. doi: 10.1007/978-1-61737-960-4_17.
Adipose stem cell preparations, either obtained as a freshly isolated so-called stromal vascular fraction (SVF) or as cells cultured to homogeneity and then referred to as adipose stem cells (ASCs), have found widespread use in a broad variety of studies on tissue engineering and regenerative medicine applications, including bone repair.For newcomers within the field, but also for established research laboratories having up to 10 years of expertise in this research area, it may be convenient to strive for, and use consensus protocols (1) for studying the osteogenic differentiation potential of ASC preparations in vitro, and (2) for osteogenic induction regimes for in vivo implementation. To assist in achieving this goal, this chapter describes various step-by-step osteogenic differentiation protocols for adipose-derived stem cell populations (SVF as well as ASCs) currently applied within our laboratory, with particular emphasis on protocols aimed at intra-operative use. The protocols describe the use of inducing compounds, including the bone morphogenetic proteins (BMPs), 1,25-dihydroxyvitamin-D3, and polyamines, as well as methods and parameters for evaluating the level of differentiation achieved.We would appreciate receiving feedback on the protocols described; this will facilitate the development of consensus protocols, which in turn will allow better comparison of data sets generated by different research groups. This continuing standardization, which might be reported on at international meetings like those of IFATS ( http://www.IFATS.org ), might be of benefit for the whole ASC research community.
脂肪干细胞制剂，无论是以新鲜分离的所谓基质血管成分（SVF）形式获得，还是培养至同质化后称为脂肪干细胞（ASC）的细胞，已在包括骨修复在内的组织工程和再生医学应用的广泛研究中得到广泛应用。对于该领域的新手，以及在该研究领域拥有长达10年专业知识的成熟研究实验室而言，遵循并使用（1）体外研究ASC制剂成骨分化潜能的共识方案，以及（2）体内实施的成骨诱导方案可能会很方便。为帮助实现这一目标，本章描述了目前我们实验室应用的针对脂肪来源干细胞群体（SVF以及ASC）的各种逐步成骨分化方案，特别强调了针对术中使用的方案。这些方案描述了诱导化合物的使用，包括骨形态发生蛋白（BMP）、1,25-二羟基维生素D3和多胺，以及评估分化程度的方法和参数。我们欢迎收到关于所描述方案的反馈；这将有助于制定共识方案，进而能更好地比较不同研究小组生成的数据集。这种持续的标准化工作（可能会在IFATS（http://www.IFATS.org）等国际会议上报告）可能会使整个ASC研究群体受益。
