Human esophageal neoplasms were studied in comparison to normal, uninvolved, and preneoplastic human esophageal epithelium for the presence of human chorionic gonadotropin (HCG), human placental lactogen (HPL), alpha-fetoprotein (AFP), carcinoembryonic antigen (CEA), and nonspecific cross-reacting antigen (NCA) using the unlabeled antibody peroxidase-antiperoxidase technique. HCG immunoreactivity was identified in 10 of 33 squamous cell carcinomas (33%), in 1 of 6 adenocarcinomas (17%), and 1 of 6 preneoplastic esophageal lesions (17%); while 9 of 33 squamous cell carcinomas (33%) and 1 of 6 adenocarcinomas (17%) contained immunoreactive AFP. Immunoreactive HPL was detected in 6 of 33 squamous cell carcinomas (20%), but in none of the adenocarcinomas. Neither AFP nor HPL immunoreactivity was identified in the 6 hyperplastic lesions which were studied. When stained with an antiserum that was able to detect both CEA and NCA, 27 of 33 squamous cell tumors (82%) and 6 of 6 adenocarcinomas (100%) showed positive immunostaining reactions. Of these, 8 squamous cell carcinomas and 1 adenocarcinoma were subsequently shown to contain only NCA immunoreactivity, while 19 squamous cell carcinomas and 5 adenocarcinomas contained both NCA and CEA immunoreactivity. NCA immunoreactivity alone was identified in 3 of 6 preneoplastic lesions and NCA and CEA immunoreactivity in 1 of 6 preneoplastic lesions. None of the markers was detected in 8 specimens of normal esophageal epithelium which were studied as controls, nor in 6 specimens of uninvolved esophageal epithelium obtained from patients with esophageal cancer. Most tumors expressed 2 or 3 markers, and some tumors were identified which expressed up to 4 of the 5 markers investigated. Only 3 tumors failed to express any of the markers studied. No association was found between the degree of tumor differentiation and presence or absence of HCG immunoreactivity. However, HPL immunoreactivity was more common in poorly differentiated squamous cell carcinomas. In contrast, immunoreactive AFP was more common in well-differentiated squamous cell carcinomas than in other tumor types. Similarly, both CEA and NCA were more frequently expressed in well-differentiated squamous cell carcinomas, adenosquamous carcinomas, and adenocarcinomas than in less differentiated tumors. Our results suggest that HCG, HPL, AFP, CEA, and NCA are tumor-associated antigens in esophageal cancer. Therefore, they could be of value in screening tests for esophageal neoplasms and could be useful in subclassification of esophageal neoplasms.

Mammary epithelium and surrounding stroma have been maintained in an explant system for 1 to 6 months and subsequently xenografted into athymic nude mice. The morphological characteristics of 26 cases of normal human mammary epithelium in long-term explant culture were described, using high-resolution light and electron microscopy. Normal human breast tissue specimens were obtained from immediate autopsy or surgical resection. The explants were cultured in Connaught Medical Research Laboratories Medium 1066 supplemented with serum, insulin, and hydrocortisone. The histotypic features of the mammary epithelium in both the central portion of the explant and the epithelial outgrowth onto the surface of the explant were described. In some cultures, the cells acquired more keratin and formed multicellular blister-like domes. Tissues from eight cases after 1 to 14 weeks in culture were xenografted in athymic nude mice and were maintained up to 924 days.

Abstract A rabbit antiserum to ovine prolactin was used with the peroxidase-antiperoxidase method to search for immunoreactive prolactin in normal and pathological human breast and prostate. We first substantiated that the heterologous antiserum could recognize human prolactin. When sections of human anterior pituitary were exposed to the anti-prolactin serum, groups of angular-shaped cells were selectively stained. The immunostaining of pituitary cells appeared specific for prolactin since both ovine and human prolactin abolished immunostaining when they were added to the anti-prolactin serum. In contrast, human growth hormone and human chorionic gonadotropin did not eliminate immunostaining. Using this anti-prolactin serum that was capable of selective recognition of prolactin-containing cells in human tissue, we stained sections of normal breast (five cases) and normal prostate (three cases). Numerous epithelial cells in these tissues were immunostained by the antiserum. The immunostaining was abolished by deletion of the prolactin antiserum and also by use of prolaction antiserum absorbed previously with human prolactin rather than the heterologous antiserum alone. Immunostaining was not eliminated by absorption of antiserum with human growth hormone or human chorionic gonadotropin. These results suggested that the immunostaining of normal breast and prostate was related to the presence of immunoreactive prolactin in epithelial cells of these organs. In prostate, immunostaining was confined to the secretory cell population; the basal cells were not stained with anti-prolactin serum. In breast, immunostaining did not appear to be confined to secretory cells but seemed to occur throughout the breast parenchyma. The immunostaining of normal breast epithelium was markedly heterogeneous with cells lacking immunoreactivity intermixed with cells having positive reactivity. This was less pronounced in normal prostate epithelium where most secretory epithelial cells were immunostained. However, marked variation in staining intensity between acini was commonly observed in normal prostate. Sixty-four % (14 of 22) of the primary prostate adenocarcinomas which were examined showed strong staining reactions with anti-prolactin serum. Two of three metastatic tumors of prostate origin had similar strong staining reactions. Most primary tumors of the prostate with strong staining reactions were poorly differentiated infiltrating adenocarcinomas (11 of 14; 79%). By contrast, 25% (three of 12) of the well-differentiated prostate adenocarcinomas and 11% (one of nine) of the benign prostate hyperplasias which were tested showed strong staining reactions when treated with anti-prolactin serum. Seven of 12 (58%) infiltrating duct carcinomas of the breast were strongly stained with anti-prolactin serum, but most breast fibroadenomas were not stained (six of eight; 75%). None of seven adenocarcinomas of the colon tested as controls were stained by anti-prolactin serum. As was the case with normal breast and prostate, absorption of anti-prolactin serum with human prolactin but not human growth hormone or human chorionic gonadotropin abolished the immunostaining of malignant breast and prostate. These results suggested that the immunostaining of breast and prostate carcinomas was related to immunoreactive prolactin. The results reported here emphasize the value of the peroxidase-antiperoxidase method in the study of hormone target organ interactions as well as in retrospective studies of pathological materials.

This chapter demonstrates that normal human mammary epithelium can be maintained in organ culture for several months. It examines that the mammary epithelium retain its secretory characteristics and epidermoid potential and) the cellular outgrowth observed is primarily myoepithelial in origin. The chapter focuses on the role of hormones in maintaining human breast epithelium in long-term organ culture system. Two approaches described are: (1) a constant regimen of breast trophic hormones present at uniform levels throughout the culture period, and (2) a cycling hormone regimen—that is, varying levels of hormones present for weekly periods in an attempt to generally mimic the in viva menstrual cycle. The chapter reviews a few reports in which normal human breast tissue from various sources was grown. These previous studies were based primarily on short-term organ culture—3–6 days. There are only three reports in which organ cultures were analyzed for over one week. Barker and others (1964) described the cultural conditions and role of insulin played in maintaining human organ cultures for up to 12 days. The chapter discusses the perspectives in terms of current applications and findings and future studies of human breast organ culture.

The chapter provides a detailed morphological, culture, and transplant study of the conditions necessary for maintaining normal human esophageal epithelium in long-term organ culture. First successful attempt to maintain normal human esophagus epithelium in a defined organ culture system for an extended period is presented in this chapter. Successful initiation of esophagus explant cultures are found to be dependent upon placing the epithelial mucosa uppermost when the tissue was placed in the petri dish. This assured that the epithelial tissue was bathed in culture media and permitted adequate diffusion of the nutrients to the cells. The proper orientation in the culture dish is also important in maintaining the normal epithelial polarity. Another important factor in the successful maintenance of esophageal epithelium culture is the inclusion of insulin and hydrocortisone in the chemically defined medium. These two hormones used in the culture system are successfully utilized singly and in combination in a variety of explant culture systems to maintain mammalian epithelia. It is possible to follow the neoplastic process in vitro using suspected organotrophic chemical carcinogens by using morphologic and ultrastructural information on normal human esophagus.