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Updated: Aug 16, 2026
Establishment of Human Epithelial Enteroids and Colonoids from Whole Tissue and Biopsy
Published on: March 6, 2015
Establishment and characterization of a colonic epithelial cell line MCE301 from transgenic mice harboring
1Molecular Genetics Research Center, Toyama Medical and Pharmaceutical University, Toyama City, Japan. ytabu@ms.toyama-mpu.ac.jp
Abstract:
We produced an immortalized colonic epithelial cell line, MCE301, using fetal mice transgenic for the temperature-sensitive simian virus 40 large T-antigen gene. MCE301 cells showed epithelial-like morphology and maintained tight connections with neighboring cells. The cells grew at a permissive temperature (33 degrees C), but the growth of the cells was significantly prevented at the nonpermissive temperature (39 degrees C). The cells expressed large T-antigen at 33 degrees C but not at 39 degrees C. MCE301 cells were not transformed, as judged by the absence of anchorage-independent growth in soft agar gel and lack of tumor formation in nude mice. Electron microscopic studies showed that the cells formed microvilli-like structures on the cell surface and junctional complexes such as tight junctions and desmosomes between the cells. The cells expressed cytosketal (acidic cytokeratins and actin), basement membrane (laminin and collagen type IV) and junctional complex proteins (ZO-1 and desmoplakin I + II), as judged by specific antibodies. Fetal bovine serum, epidermal growth factor, insulin-like growth factor and insulin significantly increased the cell growth at 33 degrees C. Moreover, MCE301 cells expressed colonic mucin Muc2 mRNA as demonstrated by reverse transcriptase-polymerase chain reaction, indicating that the cells originate from mucus-secreting cells. Alkaline phosphatase, a brush border-associated enzyme, was detected in the cells. Sodium butyrate (2 mM), an inducer of cellular differentiation, markedly elevated alkaline phosphatase activity. Thus, the present mouse colonic epithelial cell line MCE301 possessing these unique characteristics should provide a useful in vitro model of colonic epithelium.
Insights
Researchers developed MCE301, a novel mouse colonic epithelial cell line. This immortalized cell line exhibits key characteristics of normal colon cells, offering a valuable in vitro model for studying colonic epithelium.
Area of Science:
- Cell Biology
- Gastroenterology
- Animal Models
Background:
- Developing reliable in vitro models of colonic epithelium is crucial for research.
- Existing models may not fully recapitulate the complexity of native colonic tissue.
Purpose of the Study:
- To establish and characterize a new immortalized mouse colonic epithelial cell line.
- To assess the suitability of this cell line as an in vitro model for colonic epithelium research.
Main Methods:
- Immortalization of fetal mouse colonic cells using temperature-sensitive simian virus 40 large T-antigen.
- Characterization of cell morphology, growth properties, and protein expression via electron microscopy and antibody staining.
- Assessment of differentiation markers and response to growth factors and sodium butyrate.
Main Results:
- The MCE301 cell line exhibits epithelial morphology, tight cell-cell junctions, and temperature-dependent growth.
- Cells express key colonic epithelial markers including Muc2 mRNA, cytoskeletal proteins, basement membrane components, and junctional proteins.
- MCE301 cells are non-transformed, show brush border enzyme activity, and respond to differentiation inducers.
Conclusions:
- The MCE301 cell line is a well-characterized, immortalized mouse colonic epithelial model.
- It retains important structural and molecular features of native colonic epithelium.
- MCE301 serves as a valuable tool for in vitro studies of colonic epithelial biology and function.
