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The first book of its kind on marrow stromal cells, and written by leading experts in the field, this concise handbook aims to provide all that is needed to those new to this relatively young area of research. It provides a historical perspective and a summary of present knowledge of the marrow stromal lineages which are currently best understood. Detailed protocols for the isolation, culture and characterization of marrow stromal cell types from human and other animal species are given. The breadth and depth of the subject matter included makes the book equally suitable for new researchers and the more experienced investigator in this rapidly expanding field, including clinicians and bioscientists.
The stromal tissue of the bone marrow consists of a fine mesh of blood vessels held in a network of cells and extracellular matrix (ECM); it forms a continuum with the soft connective tissue adjacent to endosteal and periosteal bone surfaces and within Haversian canals. Marrow stromal tissue provides physical support and nourishment for the haemopoietic stem and progenitor cells and this led to the concept of a haemopoietic microenvironment (HME) (Dexter, Allen & Lajtha, 1977). In addition, the osteogenic potential of marrow stromal tissue has been well known for many decades (for review see Burwell, 1994).
The term ‘stromal cells’ has been used for the partially defined population of cells which make up the adherent cell layer in in vitro long-term bone marrow cultures (LTBMC) (Dexter, 1982). In this chapter, the term stromal is restricted to non-haemopoietic cells of mesenchymal origin and does not include macrophages and endothelial cells, which are also components of the adherent layer. There are three main cellular systems in the bone marrow; haemopoietic, endothelial and stromal, which are histogenetically distinct with no common precursor in the post-natal animal. The hypothesis of a marrow stromal cell system was based on analogy with the haemopoietic system (Owen, 1985); in essence, it is proposed that there are marrow stromal stem cells (MSSC) present within the bone marrow, able to generate the cell lines giving rise to the fibrous-osteogenic tissues of the skeleton and the stromal tissues of the HME (Friedenstein, 1980, 1990; Owen, 1988; Owen & Friedenstein, 1988).
Culture of marrow stromal cells cannot be covered comprehensively in a single small book. Our selection of subject matter for the different chapters was biased towards the cell lines of the marrow stromal system, which are more directly concerned with osteogenic tissues. Because of space, all authors were limited in the number of references allowed. Often a relevant review or recent paper which provides access to the previous literature has been cited as a best compromise. We apologise to those whose papers have not been quoted directly and we recognise and fully appreciate the importance of their work.
This book is dedicated to Alexander Friedenstein (1924–97), of the Gamaleya Institute, Moscow, who pioneered the subject of marrow stromal cell biology and made unrivalled contributions over many decades.