By Claire Linge (auth.), Joanna Picot (eds.)
In this moment variation of a well-liked and largely acclaimed selection of laboratory tools, a panel of major gurus have completely introduced up to date and optimized its cellphone tradition options for a large variety of human cellphone varieties proper to human sickness. each one strategy can be utilized to enquire a large spectrum of vital approaches, starting from the pathogenesis of illness, to the learn of metabolic techniques, to regulate of proliferation and differentiation. New to this variation are chapters on fibroblasts, Schwann cells, gastric and colonic epithelial cells, and parathyroid cells. The protocols stick to the profitable tools in Molecular medication™ sequence layout, each one supplying step by step laboratory directions, an advent outlining the primary in the back of the method, lists of the mandatory apparatus and reagents, and pointers on troubleshooting and keeping off identified pitfalls.
Wide-ranging and hugely sensible, Human mobile tradition, moment variation, offers amateur and skilled researchers alike with a close, step by step consultant to winning tradition human cells today.
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J. Biol. Chem. 267, 24,675–24,680. 18. 18 Hirobe, T. (2001) Endothelins are involved in regulating the proliferation and differentiation of mouse epidermal melanocytes in serum-free primary culture. J. Invest. Dermatol. Symp. Proc. 6, 25–31. 19. , et al. (1998) Endothelin-1 is a paracrine growth factor that modulates melanogenesis of human melanocytes and participates in their responses to ultraviolet radiation. Cell Growth Differ. 9, 575–584. 20. , and Le Douarin, N. M. (1996) Endothelin-3 promotes neural crest cell proliferation and mediates a vast increase in melanocyte number in culture.
Phenotypic Stability in Culture As a matter of routine, we perform our studies on cells at first passage. Other investigators have studied the effects of repeated subculture on the phenotypic stability of HBDCs and found that they lose their osteoblast-like characteristics. In practical terms, this presents real difficulties, because it is often desirable to obtain large numbers of HBDCs from a single donor. As an alternative to repeated subculture, trabecular explants can be replated at the end of primary culture into a new flask (see Fig.
4. To screen for potential therapeutic agents. 5. To test and develop new biomaterials. 6. To use cell therapy and tissue engineering for bone transplantation. The technique for culturing primary human osteoblasts have been reviewed widely over the past few years (6–8); however, in this rapidly moving field there have been many new developments. The aim of this chapter is threefold: to describe the basic techniques of bone cell culture (both primary and clonal); to update researchers on new developments in the methodology used to characterize these cells; and to highlight the investigative value of bone cell cultures, including new techniques in which progenitor cells, particularly mesenchymal stem cells, are used to assess the effects of environmental cues on osteoblast differentiation.