In vitro culture of embryonic mouse intestinal epithelium: cell differentiation and introduction of reporter genes

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• 作者：Jonathan M Quinlan (1)
  Wei-Yuan Yu (1) (2)
  Mark A Hornsey (1)
  David Tosh (1)
  Jonathan M W Slack (1)
  
• 刊名：BMC Developmental Biology
• 出版年：2006
• 出版时间：December 2006
• 年：2006
• 卷：6
• 期：1
• 全文大小：1967KB
• 参考文献：1. Slack JMW: Development of endodermal organs. / Essential Developmental Biology / 2 EditionBlackwell Scientific 2006, 249鈥?63.
  2. Mathan M, Moxey PC, Trier JS: Morphogenesis of fetal rat duodenal villi. / Am J Anat 1976, 146:73鈥?2. CrossRef
  3. Potten CS, Loeffler M: Stem cells: attributes, cycles, spirals, pitfalls and uncertainties. Lessons for and from the crypt. / Development 1990, 110:1001鈥?020.
  4. Potten CS: Stem cells in gastrointestinal epithelium: numbers, characteristics and death. / Phil Trans R Soc London Ser B 1998, 353:821鈥?30. CrossRef
  5. Sancho E, Batlle E, Clevers H: Signaling pathways in intestinal development and cancer. / Ann Rev Cell Dev Biol 2004, 20:695鈥?23. CrossRef
  6. Real FX, Xu M, Vila MR, de Bolos C: Intestinal brush鈥揵order associated enzymes: co鈥搊rdinated expression in colorectal cancer. / Int J Cancer 1992, 51:173鈥?81. CrossRef
  7. Hocker M, Wiedenmann B: Molecular mechanisms of enteroendocrine differentiation. / Ann NY Acad Sci 1998, 859:160鈥?74. CrossRef
  8. Roth KA, Gordon JL: Spatial differentiation of the intestinal epithelium: analysis of enteroendocrine cells containing immunoreactivity serotonin, secretin, and substance P in normal and transgenic mice. / Proc Natl Acad Sci USA 1990, 87:6408鈥?412. CrossRef
  9. Porter EM, Bevins CL, Ghosh D, Ganz T: The multifaceted Paneth cell. / Cell Mol Life Sci 2002, 59:156鈥?70. CrossRef
  10. Cheng H, Leblond CP: Origin, differentiation, and renewal of the four main epithelial cell types in the mouse small intestine. V Unitarian Theory of the origin of the four epithelial cell types. / Am J Anat 1974, 141:537鈥?62. CrossRef
  11. Wright NA: Epithelial stem cell repertoire in the gut: clues to the origin of cell lineages, proliferative units and cancer. / Int J Exp Pathol 2000, 81:117鈥?43. CrossRef
  12. Whitehead RH, Demmler K, Rockmann SP, Watson NK: Clonogenic growth of epithelial cells from normal colonic mucosa from both mice and humans. / Gastroenterology 1999, 117:858鈥?65. CrossRef
  13. Hall PA, Coates PJ, Ansari B, Hopwood D: Regulation of cell number in the mammalian gastrointestinal tract: the importance of apoptosis. / J Cell Sci 1994, 107:3569鈥?577.
  14. Strater J, Koretz K, Gunthert AR, Moller P: In situ detection of enterocytic apoptosis in normal colonic mucosa and in familial adenomatous polyposis. / Gut 1995, 37:819鈥?25. CrossRef
  15. Tait IS, Evans GS, Kedinger M, Flint N, Potten CS, Campbell FC: Progressive morphogenesis in vivo after transplantation of cultured small bowel epithelium. / Cell Transplant 1994, 3:33鈥?0.
  16. Simon鈥揂ssmann P, Kedinger M: Heterotypic cellular cooperation in gut morphogenesis and differentiation. / Semin Cell Biol 1993, 4:221鈥?30. CrossRef
  17. Hearn CJ, Young HM, Ciampoli D, Lomax AE, Newgreen D: Catenary cultures of embryonic gastrointestinal tract support organ morphogenesis, motility, neural crest cell migration, and cell differentiation. / Dev Dyn 1999, 214:239鈥?47. CrossRef
  18. Abud HE, Lock P, Heath JK: Efficient gene transfer into the epithelial cell layer of embryonic mouse intestine using low鈥搗oltage electroporation. / Gastroenterology 2004, 126:1779鈥?787. CrossRef
  19. Abud HE, Watson N, Heath JK: Growth of intestinal epithelium in organ culture is dependent on EGF signaling. / Exp Cell Res 2005, 303:252鈥?62. CrossRef
  20. Percival AC, Slack JMW: Analysis of pancreatic development using a cell lineage label. / Exp Cell Res 1999, 247:123鈥?32. CrossRef
  21. Horb LD, Slack JMW: Role of cell division in branching morphogenesis and differentiation of the embryonic pancreas. / Int J Dev Biol 2000, 44:791鈥?96.
  22. Shen C鈥揘, Seckl JR, Slack JMW, Tosh D: Glucocorticoids suppress beta cell development and induce hepatic metaplasia in embryonic pancreas. / Biochem J 2003, 375:41鈥?0. CrossRef
  23. Yu W鈥揧, Slack JMW, Tosh D: Conversion of columnar to stratified squamous epithelium in the developing mouse oesophagus. / Dev Biol 2005, 284:157鈥?70. CrossRef
  24. Silberg DG, Swain GP, Suh ER, Traber PG: Cdx1 and Cdx2 expression during intestinal development. / Gastroenterology 2000, 119:961鈥?71. CrossRef
  25. Yuasa Y: Control of gut differentiation and intestinal鈥搕ype gastric carcinogenesis. / Nature Rev Cancer 2003, 3:592鈥?00. CrossRef
  26. Owens DW, Lane EB: The quest for the function of simple epithelial keratins. / Bioessays 2003, 25:748鈥?58. CrossRef
  27. Stappenbeck TS, Gordon JI: Rac1 mutations produce aberrant epithelial differentiation in the developing and adult mouse small intestine. / Development 2000, 127:2629鈥?642.
  28. Bry L, Falk P, Huttner K, Ouellette A, Midvedt T, Gordon JI: Paneth cell differentiation in the developing intestine of normal and transgenic mice. / Proc Natl Acad Sci USA 1994, 91:10335鈥?0339. CrossRef
  29. Zhong W, Mirkovitch J, Darnell JE Jr: Tissue鈥搒pecific regulation of mouse hepatocyte nuclear factor 4 expression. / Mol Cell Biol 1994, 14:7276鈥?284.
  30. Grapin鈥揃otton A, Majithia AR, Melton DA: Key events of pancreas formation are triggered in gut endoderm by ectopic expression of pancreatic regulatory genes. / Genes Dev 2001, 15:444鈥?54. CrossRef
  31. Itasaki N, Bel鈥揤islar S, Krumlauf R: "Shocking" developments in chick embryology:electroporation and in ovo gene expression. / Nature Cell Biology 1999, 1:E203鈥揈207. CrossRef
  32. Saito T, Nakatsuji N: Efficient gene transfer into embryonic mouse brain using in vivo electroporation. / Dev Biol 2001, 240:237鈥?46. CrossRef
  33. Shen C鈥揘, Slack JMW, Tosh D: Molecular basis of transdifferentiation of pancreas to liver. / Nature Cell Biology 2000, 2:879鈥?87. CrossRef
  34. Tosh D, Shen C鈥揘, Slack JMW: Differentiated properties of hepatocytes induced from pancreatic cells. / Hepatology 2002, 36:534鈥?43. CrossRef
  35. Graham FL, Smiley J, Russell WC, Nairn R: Characteristics of a human cell line transformed by DNA from human adenovirus type 5. / J Gen Virol 1977, 36:59鈥?4. CrossRef
  36. Graham FL, Prevec L: Adenovirus鈥揵ased expression vectors and recombinant vaccines. / Biotechnology 1992, 20:363鈥?90.
  37. Cohen CJ, Shieh JTC, Pickles RJ, Okegawa T, Hsieh JT, Bergelson JM: The coxsackievirus and adenovirus receptor is a transmembrane component of the tight junction. / Proc Natl Acad Sci USA 2001, 98:T15191鈥?5196. CrossRef
  38. Black BL, Moog F: Alkaline phosphatase and maltase activity in the embryonic chick intestine in culture. / Dev Biol 1978, 66:232鈥?49. CrossRef
  39. Bleich HL, Boro ES, Trier JS: Organ鈥揷ulture methods in the study of gastrointestinal鈥搈ucosal function and development. / N Engl J Med 1976, 295:150鈥?55. CrossRef
  40. DeRitis G, Falchuk ZM, Trier JS: Differentiation and maturation of cultured fetal rat jejunum. / Dev Biol 1975, 45:304鈥?17. CrossRef
  
• 作者单位：Jonathan M Quinlan (1)
  Wei-Yuan Yu (1) (2)
  Mark A Hornsey (1)
  David Tosh (1)
  Jonathan M W Slack (1)
  
  1. Centre for Regenerative Medicine, Department of Biology & Biochemistry, University of Bath, Bath, BA2 7AY, UK
  2. Department of Craniofacial Development, King's College London, Floor 28, Guy's Hospital, London Bridge, London, SE1 9RT, UK
  

文摘

Background Study of the normal development of the intestinal epithelium has been hampered by a lack of suitable model systems, in particular ones that enable the introduction of exogenous genes. Production of such a system would advance our understanding of normal epithelial development and help to shed light on the pathogenesis of intestinal neoplasia. The criteria for a reliable culture system include the ability to perform real time observations and manipulations in vitro, the preparation of wholemounts for immunostaining and the potential for introducing genes. Results The new culture system involves growing mouse embryo intestinal explants on fibronectin-coated coverslips in basal Eagle's medium+20% fetal bovine serum. Initially the cultures maintain expression of the intestinal transcription factor Cdx2 together with columnar epithelial (cytokeratin 8) and mesenchymal (smooth muscle actin) markers. Over a few days of culture, differentiation markers appear characteristic of absorptive epithelium (sucrase-isomaltase), goblet cells (Periodic Acid Schiff positive), enteroendocrine cells (chromogranin A) and Paneth cells (lysozyme). Three different approaches were tested to express genes in the developing cultures: transfection, electroporation and adenoviral infection. All could introduce genes into the mesenchyme, but only to a small extent into the epithelium. However the efficiency of adenovirus infection can be greatly improved by a limited enzyme digestion, which makes accessible the lateral faces of cells bearing the Coxsackie and Adenovirus Receptor. This enables reliable delivery of genes into epithelial cells. Conclusion We describe a new in vitro culture system for the small intestine of the mouse embryo that recapitulates its normal development. The system both provides a model for studying normal development of the intestinal epithelium and also allows for the manipulation of gene expression. The explants can be cultured for up to two weeks, they form the full repertoire of intestinal epithelial cell types (enterocytes, goblet cells, Paneth cells and enteroendocrine cells) and the method for gene introduction into the epithelium is efficient and reliable.