FIGURE 22.1 Isolation of Organoid Structures from Breast. (a) Flowchart of isolation of different fractions by selective sedimentation and centrifugation following collagenase digestion. (b) A TDLUlike structure derived from a Muc-/ESA+ /CD29hi D920 cell grown in collagen I/laminin-1 three- dimensional culture Labarge et al., The structure was stained for keratin-14 (red) and keratin-8 (green) expression, and imaged with a confocal microscope. The presented image is a reconstruction of several optical slices to give to appearance of three dimentions. There is a lumen indicated by a star. Scale bar = 50 μm. (See also Plate 26; Reproduced from LaBarge et al., 2007.)

FIGURE 22.2 HepaRG Cells. Phase contrast of HepaRG living cells. (a) Four-week-old differentiated culture maintained for 2 weeks in presence of 2% DMSO. Note the mixed population with highly differentiated hepatocytes colonies bordered by flat biliary cells. (b) Two-day-old culture of selected HepaRG hepatocytes maintained in presence of 2% DMSO. Note the purity of the population and the hepatocyte organization in typical hepatic trabeculae.

FIGURE 22.3 Vascular Endothelial Cells. (a) View of blood vessels in human umbilical cord. Umbilical arteries (black arrows); umbilical vein (white arrow) with Luer adaptor inserted (see also Plate 8e). (b) View of human aortic endothelial cell monolayer under phase contrast microscope. 10x magnification.

FIGURE 22.4 Olfactory Bulb Dissection. Schematic diagram of the isolation of the olfactory bulbs from the brain of newborn rats. (Courtesy of Susan Barnett.)

FIGURE 22.5 Melanocyte Cultures. (a) Culture of Caucasian newborn foreskin-derived melanocytes 1 week after inoculation. Note the multiple keratinocyte colonies with central stratification and tightly apposed epithelial cells at the periphery. The melanocytes are the relatively small, dark dendritic cells, most of them in contact with the keratinocyte colonies by means of dendritic projections (seen via phase contrast, 100× magnification). (b) Ten-day-old primary cultures of Caucasian newborn epidermal melanocytes in medium lacking TPA. Many cells display branching dendrites, and other cells display bipolar to polygonal morphology (seen via phase contrast, 40× magnification). (c) Secondary culture of TPA-treated epidermal melanocytes derived from African-American newborn foreskin. Note the dendritic morphology of the cells and their slender spindle shape. In contrast to the melanocytes derived from Caucasian newborn foreskin, these melanocytes display a high level of melanin granules (seen via phase contrast, 320× magnification). (See also Plate 8d; photographs courtesy of Hee-Young Park.)