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12.6. CONCLUSIONS It is neither possible nor ethical to perform genetic engineering experiments in humans; thus development of human mammary cell culture techniques has been crucial and indeed necessary to locate, isolate, and characterize putative human mammary stem cells. Clearly, the models will need to be improved to even more accurately re ect the microenvironment so that existing and yet-to-be developed powerful culture techniques can be brought to bear on a number of new questions: How do stem cells integrate signals that control their functions and behavior What role does the microenvironment (other cells, hormones, growth factors, ECM molecules in short the niche) play in normal mammary gland morphogenesis and in the initiation and propagation of tumorigenesis How do stem cells relate to cancer What is the cellular etiology of different types of breast cancers It has been observed that histological subtypes in cancers emerge that appear lineage restricted, suggesting that stem and progenitor cells might lie at the cancer s origin. This scenario has been documented in the evolution of two distinct leukemias: Chronic myeloid leukemia is derived from hematopoietic stem cells, and acute myeloid leukemia is derived from committed granulocyte-macrophage progenitors [Daley, 2004; Jamieson et al., 2004]. Stemlike cells, isolated from a breast tumor, could be serially transplanted as a tumor in nude mice, but whether these were directly related to normal mammary stem or progenitor cells is not known [Al-Hajj et al., 2003]. It was additionally hypothesized that the stem cell microenvironment may play an important initiating or promoting role in tumorigenesis [reviewed in Bissell and Labarge, 2005]. Now that a hierarchy among mammary stem and progenitor cells has been identi ed for rodents and, more recently, for humans [Villadsen et al., 2007], as well as their respective locations within the mammary gland, a detailed analysis of their transformed phenotypes and what roles their microenvironments play in fostering speci c behaviors should take place. One particular challenge will be providing unequivocal proof that one normal stem cell does give rise to a mammary gland, and that the same cell, if transformed, gives rise to a tumor in vivo. Because 3D culture environments can model form and functions as complex as TDLU formation from single cells, they may also provide a malleable proving ground to demonstrate such a relationship. 12.7. SOURCES OF MATERIALS Item Anti-mouse IgG magnetic beads Ascorbic acid B27 supplement Bovine serum albumin (fraction V) Cholera toxin Collagenase Culture asks, 25 cm2 Dibutyryl cAMP Supplier Miltenyi Biotech Sigma Invitrogen (GIBCO) Sigma Sigma Sigma Nunc Sigma NOTE
Excluding page elements from error checking
Summary Use the Flickr API Add geodata to your Flickr photos Embed Flickr photos into a Google Map
NOTE
See 9 for implementation notes for a filter relationship using the essentials of this technique.
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