Abstract
Human esophageal epithelium obtained from intermediate autopsies (<12 h) was maintained as cell and explant cultures. In order to develop a serum-free, defined media culture model, several medias and additives were evaluated. The viability and differentiation of the epithelial cells cultured with serum-free, Keratinocyte Growth Media (KGM, Clonetics Co., San Diego, CA) was improved over that of esophageal cells and explants cultured in either serum-supplemented CMRL 1066 (OCM), serum-free additive-supplemented CMRL 1066, or cimetidine-supplemented CMRL 1066. The KGM component EGF was determined to be trophic for esophagus cells on the basis of findings of increased 3H-TdR tabelling in KGM cultures when compared to control cells grown in KGM without EGF (KBM). The morphologic pattern of the cytoskeletal proteins actin, keratin, and vimentin were characterized in isolated cell populations. The intermediate filaments, keratin, and vimentin were co-expressed in these epithelial cells. Esophageal explant viability, differentiation, and outgrowth from 15 cases were also evaluated in dishes coated with basement membrane associated proteins. Explants cultured in these dishes were equally well-preserved and differentiated. There were no significant differences in the explant histology when there was protein coating of the culture dishes, although one case showed improved outgrowth with laminin coating. A main advantage for using this culture system is that the same medium (KGM) can be used for both the culture of explants and isolated epithelial cells. Future applications of this model include determining: (1) the effect different concentrations of EGF and calcium in the media will have on esophageal proliferation and differentiation, and (2) the role of different basement membrane associated proteins on the plating efficiency of either isolated or outgrowth epithelial esophageal cells.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Arsenault P and Menard D (1988) Autoradiographic localization of [3H]-thymidine incorporation in developing human esophagus. Anat. Rec. 220: 313–317.
Banks-Schlegel SP (1985) Isolation, cultivation, and characterization of normal human esophageal epithelial cells. J. Tis. Cult. Methods 9: 95–105.
Banks-Schlegel SP and Quintero J (1986) Growth and differentiation of human esophageal carcinoma cell lines. Cancer Res. 46: 250–258.
Barrett LA, McDowell EM, Harris CC and Trump BF (1977) Studies on the pathogenesis of ischemic cell injury. IV. Reversal of ischemic cell injury in hamster trachea and human bronchus by explant culture. Beitr. Path. 161: 109–121.
Blay J and Brown KD (1986) Contradistinctive growth response of cultured rat intestinal epithelial cells to epidermal growth factor depending on cell population density. J. Cell. Physiol 129: 343–346.
Blom H (1983) Cimetidine and parietal cell regeneration in experimental wounds in rat gastric mucosa. Scand. J. Gastroenterol. 18: 853–857.
Chopra DP, Siddiqui KM and Cooney RA (1987) Effects of insulin, transferrin, cholera toxin, and epidermal growth factor on growth and morphology of human fetal normal colon epithelial cells. Gastroenterology 92: 891–904.
Cottrell JR and Resau JH (1985) A method to determine the viability of explant tissue in organ culture. J. Tis. Cult. Methods 9: 191–192.
Hennings H, Michael D, Cheng C, Steinert P, Holbrook K and Yuspa SH (1980) Calcium regulation of growth and differentiation of mouse epidermal cells in culture. Cell 19: 245–254.
Keenan KP, Combs J and McDowell EM (1982) Regeneration of hamster tracheal epithelium after mechanical injury. IV. Histochemical, immunocytochemical and ultrastructural studies. Virchows Arch. [B] 43: 213–240.
Konturek SJ (1988a) Role of epidermal growth factor in gastroprotection and ulcer healing. Scand. J. Gastroenterol. 23: 129–133.
Konturek SJ, Dembinski A, Warzecha Z, Brzozowski T and Gregory H (1988b) Role of epidermal growth factor in healing of chronic gastroduodenal ulcers in rats. Gastroenterology 94: 1300–1307.
Lechner J and LaVeck M (1985) A serum-free method for culturing normal human bronchial epithelial cells at clonal density. J. Tis. Cult. Methods 9: 43–52.
Lille JH, MacCallum DK and Jepsen A (1988) Growth of stratified squamous epithelium on reconstituted extracellular matrices: Long-term culture. J. Invest. Dermatol. 90: 100–109.
Lipkin M, Friedman E, Winawer SJ and Mewmark H (1989) Colonic epithelial cell proliferation in responders and nonresponders to supplemental dietary calcium. Cancer Res. 49: 248–254.
Menard D and Arsenault P (1987) Maturation of human fetal esophagus maintained in organ culture. Anat. Rec. 217: 348–354.
Mohamed S, Nadijeka M and Hanson U (1986) Wound healing properties of cimetidine in vitro. Drug Intell. Clin. Pharm. 20: 973–975.
Moyer MP (1983) Rapid Communications. Culture of human gastrointestinal epithelial cells. Proc. Soc. Exp. Biol. Med. 174: 12–15.
Orlando RC, Turjman NA, Tobey NA, Scheneiner VJ and Powell DW (1987) Mucosal protection by sucralfate and its components in acid-exposed rabbit esophagus. Gastroenterology 93: 32–61.
Peehl D and Ham RG (1980) Growth and differentiation of human keratinocytes without a feeder layer or conditioned medium. In Vitro 16: 516–525.
Resau JH, He AG and Jones RT (1983a) Establishment of adult Syrian golden hamster duodenal epithelial cultures from organ explants. J. Tis. Cult. Methods 8: 181–184.
Resau JH, Hudson EA and Jones RT (1983b) Organ explant culture of Syrian golden hamster pancreas. In Vitro 19: 315–325.
Resau JH, Sakamoto K, Cottrell JR and Newkirk C (1986) Explant culture of hamster alimentary tract epithelium. Cell. Biol. Toxicol. 2: 401–415.
Resau JH, Cottrell JR, Elliget KA and Hudson EA (1987) Cell injury and regeneration of human epithelium in organ culture. Cell Biol. Toxicol. 3: 441–458.
Resau JH, Phelps PC, He A, Anthony RA and Jones RT (1988) Long-term culture of hamster duodenal explants and cells. Digestion 41: 9–21.
Shipley GD and Pittelkow MR (1987) Control of growth and differentiation in vitro of human keratinocytes cultured in serum-free medium. Arch. Dermatol. 123: 1541a-1544a
Thomas de la Vega J, Banner B, Haklin M, Saxon A and Roseman D (1983) Effect of cimetidine on intestinal adaptation following massive resection of small intestine. Surg. Gynecol. Obstet. 156: 41–50.
Trump BF, Valagorsky JM, Jones RT, Mergner WJ, Garcia JH and Cowley RA (1975) The application of electron microscopy and cellular biochemistry to the autopsy. Observation on cellular change in human shock. Hum. Pathol. 6: 499–516.
Vocci MJ, Combs JW, Hillman EA, Resau JH and Trump BF (1983) The cell kinetics of the adaption of the esophagus to organ culture. In vitro 19: 881–891.
Author information
Authors and Affiliations
Additional information
This is publication #2544 from the Pathobiology Laboratory.
Rights and permissions
About this article
Cite this article
Resau, J.H., Phelps, P.C., Zhu, Sm. et al. Long-term culture of human esophageal explants and cells. Cytotechnology 3, 61–73 (1990). https://doi.org/10.1007/BF00365267
Received:
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/BF00365267