Human esophageal, tracheal, and pancreatic ductal fragments were collected at autopsy after a postmortem interval of 12 hours or less and maintained in explant organ culture for 30 days. The viability and growth of the explants was assessed by morphology, LDH enzyme release, and cellular outgrowth. The viability and growth of the bronchial explant epithelium was directly related to the postmortem interval. Esophageal epithelial regeneration followed the desquamation of the superficial cell layers. Pancreatic epithelia appeared to grow more slowly and with less outgrowth than the other tissues. Epithelial cell growth along the explant surface and onto the culture dish appeared to proceed through the well-characterized process that follows cell injury, i.e., flattening, migration, replication, and differentiation. Thus, sufficient numbers of viable epithelial cells capable of regeneration were present in routine autopsy epithelium, but there was considerable variation from tissue to tissue and case to case. The most effective and accurate approach to follow when evaluating and predicting the growth and viability of these explants is by using a combination of morphologic, enzymatic and biologic assays. Errors in the interpretation of viability are possible when only one assay method is utilized. These tissues grown in explant organ culture are suitable for studies on the mechanism and response of epithelia to cell injury, recovery and wound healing.
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Abbreviations
- 4F-1G:
-
4% formaldehyde, 1% glutaraldehyde
- HIFBS:
-
heat inactivated fetal bovine serum
- IA:
-
immediate autopsy
- LDH:
-
lactate dehydrogenase
- OsO4 :
-
osmium tetroxide
- RA:
-
routine autopsy
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Resau, J.H., Cottrell, J.R., Elligett, K.A. et al. Cell injury and regeneration of human epithelium in organ culture. Cell Biol Toxicol 3, 441–458 (1987). https://doi.org/10.1007/BF00119916
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DOI: https://doi.org/10.1007/BF00119916