Selective growth of some rodent epithelial cells in a medium containing citrulline.

Sun, N. C.; Sun, Cheng‐He; Tennant, R. W.; Hsie, Abraham W.

doi:10.1073/pnas.76.4.1819

Cited by 21 publications

(8 citation statements)

References 15 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Day-12 and -34 cells, on the other hand, not only formed monolayers but also exhibited significant trans -differentiation into fibroblast-like mesenchymal cells within 6–12 days post plating (Figure 2b). These changes were not driven by contamination from the mesenchymal cells during the isolation process, as the purified crypts (Supplementary Figures 1D and E) were negative for α-smooth muscle actin expression either in uninfected or in 6–34-day post-infected crypt cellular extracts (Supplementary Figure 1F), whereas cells isolated from uninfected mouse distal colon not only failed to grow beyond 1–2 weeks in culture conditions that included D-valine 40 or citrulline, 41 but never changed their phenotype (data not shown). When the cells in monolayers were stained for markers of EMT, they stained positive for vimentin but negative for E-cadherin, suggesting an EMT-like process in vitro (Figure 2d).…”

Section: Resultsmentioning

confidence: 98%

Utility of a bacterial infection model to study epithelial–mesenchymal transition, mesenchymal–epithelial transition or tumorigenesis

et al. 2013

View full text Add to dashboard Cite

DCLK1 and Lgr5 have recently been identified as markers of quiescent and cycling stem cells in the small intestinal crypts, respectively. Epithelial–mesenchymal transition (EMT) is a key development program that is often activated during cancer invasion and metastasis, and also imparts a self-renewal capability to disseminating cancer cells. Utilizing the Citrobacter rodentium (CR)-induced transmissible murine colonic hyperplasia (TMCH) model, we observed a relative decrease in DCLK1 expression in the colonic crypts, with significant shift towards stromal staining at peak (12 days post infection) hyperplasia, whereas staining for Lgr5 and Msi-1 increased several fold. When hyperplasia was regressing (days 20–34), an expansion of DCLK1 +ve cells in the CR-infected crypts compared with that seen in uninfected control was recorded. Purified colonic crypt cells exhibiting epigenetic modulation of the transforming growth factor-β (TGFβ), Wnt and Notch pathways on 12 or 34 days post infection formed monolayers in vitro, and underwent trans-differentiation into fibroblast-like cells that stained positive for vimentin, fibronectin and DCLK1. These cells when trypsinized and regrown in soft agar, formed colonospheres/organoids that developed into crypt-like structures (colonoids) in Matrigel and stained positive for DCLK1. Mice exhibiting 12 or 34 days of TMCH were given azoxymethane once for 8 h (Gp1) or weekly for 3 weeks (Gp2), and subjected to crypt isolation. Crypt cells from Gp1 animals formed monolayers as well as colonospheres in soft agar and nodules/tumors in nude mice. Crypt cells isolated from Gp2 animals failed to form the monolayers, but developed into colonospheres in soft agar and nodules/tumors in nude mice. Thus, both hyperplasia and increased presence of DCLK1 +ve cells promote cellular transformation in response to a second hit. The TMCH model, therefore, provides an excellent template to study how alterations in intestinal stem cells promote trans-differentiation, crypt regeneration or colon carcinogenesis following bacterial infection.

show abstract

Section: Resultsmentioning

confidence: 98%

Utility of a bacterial infection model to study epithelial–mesenchymal transition, mesenchymal–epithelial transition or tumorigenesis

et al. 2013

View full text Add to dashboard Cite

show abstract

“…This finding did not receive the recognition it may have deserved as it was widely known that most mammalian cells do not require arginine for growth in tissue culture because media that contain citrulline in place of arginine can most often support normal growth. 6,10 In 1979, Sun et al 19 demonstrated definitively that some tumor cells are unable to utilize citrulline for growth. However, most of these investigators concluded that Mycoplasma contamination of tissue culture should be avoided.…”

Section: Discussionmentioning

confidence: 99%

Incidence and distribution of argininosuccinate synthetase deficiency in human cancers

Dillon

Prieto

Curley

et al. 2004

Cancer

270

229

View full text Add to dashboard Cite

BACKGROUND Argininosuccinate synthetase (ASS) was the first of two enzymes to convert citrulline to arginine. This pathway allowed cells to synthesize arginine from citrulline, making this amino acid nonessential for the growth of most mammalian cells. Previous studies demonstrated that several human tumor cell lines were auxotrophic for arginine due to an inability to express ASS. Selective elimination of arginine from the circulation of animals with these tumors is a potentially effective anticancer treatment. The purpose of these experiments was to determine the frequency of ASS deficiency and arginine auxotrophy in a variety of human malignant tumors. METHODS The authors analyzed the expression of ASS by immunohistochemistry with a monoclonal antibody in a variety of human tumor biopsies. They found that the incidence of ASS deficiency varied greatly with the tumor type and tissue of origin. RESULTS Melanoma, hepatocellular carcinoma, and prostate carcinoma were most frequently deficient in ASS. Some human cancers were almost always positive for ASS (e.g., lung and colon carcinomas). However, other human cancers, including sarcomas, invasive breast carcinoma, and renal cell carcinoma, also were sometimes ASS deficient. CONCLUSIONS These data indicated that immunohistochemical detection of ASS may prove an effective means for determining ASS deficiency in malignant human tumors and for identifying patients most likely to respond to arginine deprivation therapy. Based on these results, human clinical trials using arginine‐degrading enzyme therapy to treat patients with advanced melanoma or hepatocellular carcinoma have been initiated. Cancer 2004;100:826–33. © 2004 American Cancer Society.

show abstract

“…There have been numerous but inconclusive attempts to solve this problem by the use of selective media that inhibit fibroblast growth . These include media in which L-valine is replaced by D-valine (4), media in which arginine is replaced by citrulline (21), and media lacking tyrosine (2) proach to overcoming the problem of fibroblast overgrowth . Although colostrum, in the absence of serum, supports the growth of such epithelial cells as the MDCK cell line, milk obtained as early as a week after the birth of a calf is completely inactive.…”

Section: Discussionmentioning

confidence: 99%

Bovine colostrum supports the serum-free proliferation of epithelial cells but not of fibroblasts in long-term culture

Klagsbrun¹

1980

The Journal of Cell Biology

View full text Add to dashboard Cite

Selective growth of some rodent epithelial cells in a medium containing citrulline.

Abstract: We have defined a medium (called Sun

Cited by 21 publications

References 15 publications

Utility of a bacterial infection model to study epithelial–mesenchymal transition, mesenchymal–epithelial transition or tumorigenesis

Utility of a bacterial infection model to study epithelial–mesenchymal transition, mesenchymal–epithelial transition or tumorigenesis

Incidence and distribution of argininosuccinate synthetase deficiency in human cancers

Bovine colostrum supports the serum-free proliferation of epithelial cells but not of fibroblasts in long-term culture

Contact Info

Product

Resources

About