By Claire Linge (auth.), Joanna Picot (eds.)
In this moment variation of a well-liked and greatly acclaimed number of laboratory equipment, a panel of top specialists have completely introduced updated and optimized its mobile tradition suggestions for a wide variety of human telephone varieties suitable to human illness. every one method can be utilized to enquire a large spectrum of significant tactics, starting from the pathogenesis of sickness, to the learn of metabolic strategies, to manage of proliferation and differentiation. New to this version are chapters on fibroblasts, Schwann cells, gastric and colonic epithelial cells, and parathyroid cells. The protocols stick with the winning equipment in Molecular medication™ sequence structure, each one providing step by step laboratory directions, an advent outlining the primary in the back of the approach, lists of the mandatory gear and reagents, and tips about troubleshooting and keeping off recognized pitfalls.
Wide-ranging and hugely sensible, Human phone tradition, moment version, presents beginner and skilled researchers alike with a close, step by step advisor to winning tradition human cells today.
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Additional info for Human Cell Culture Protocols
Basal activity is initially low, but increases with increasing cell density. Treatment with 1,25(OH)2D3 increases alkaline phosphatase activity. The most specific phenotypic marker is osteocalcin. This is a protein of Mr 5800 containing residues of the vitamin K–dependent amino acid γ-carboxyglutamic acid. In humans, its synthesis is restricted to mature cells of the osteoblast lineage. It is an excellent late stage marker for cells of this series despite the fact that its precise function in bone has yet to be established.
Addition of L-ascorbic acid (50 μg/mL–25 μM) (see Note 1) to HBDCs in secondary culture (E1P1) increases proliferation, produces a sustained increase in the steady-state levels of α1 (I)-procollagen mRNA, and dramatically increases the secretion of type I collagen. There is also an increase in noncollagenous protein synthesis, most importantly bone sialoprotein and osteocalcin, and increased deposition of matrix. 5. Passaging Cells Cultured in the Continuous Presence of Ascorbate Because of their synthesis and secretion of an extensive collagen-rich extracellular matrix, HBDCs cultured in the continuous presence of ascorbate cannot be subcultured using trypsin-EDTA alone.
J. (1986) Folliclestimulating hormone enhances somatomedin C binding to cultured rat granulosa cells. J. Biol. Chem. 261, 3923–3926. 32. 32 Gilchrest, B. , Vrabel, M. , and Szabo, G. (1984) Selective cultivation of human melanocytes from newborn and adult epidermis. J. Invest. Dermatol. 83, 370–376. 33. Medawar, P. B. (1941) Sheets of pure epidermal epithelium from human skin. Nature 148, 783. 34. Niedel, J. E. and Blackshear, P. J. (1986) Protein kinase C, in Phosphoinositides and Receptor Mechanisms (Putney, J.
Human Cell Culture Protocols by Claire Linge (auth.), Joanna Picot (eds.)