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| Status |
Public on Oct 07, 2009 |
| Title |
Immortalized colonic epithelial progenitor cells express stem cell markers and differentiate in vitro |
| Organism |
Homo sapiens |
| Experiment type |
Expression profiling by array
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| Summary |
Immortalized colonic epithelial progenitor cells derived from normal human colon biopsies express stem cell markers and differentiate in vitro
The sustained growth of normal non-transformed human colonic epithelial cells has historically been exceptionally challenging. We here report the successful long-term propagation of colon biopsy derived cells expressing both epithelial and mesenchymal features, intestinal stem cell markers, and demonstrating multilineage epithelial differentiation capability. Cells isolated from the biopsies of two patients undergoing routine screening colonoscopy have been successfully passaged in vitro, subsequently immortalized with non-oncogenic proteins (cyclin dependent kinase 4 [CDK4] and the catalytic component of human telomerase [hTERT]), and maintained for well over a year in continuous cell culture. Immunofluorescence experiments reveal the immortalized populations of cells significantly express the progenitor epithelial cell marker mucin-1 and the colon epithelial specific marker A33. In addition, staining shows subsets of cells express chromogranin A, mucin-2, and dipeptyl peptidase 4, corresponding to neuroendocrine, mucus secreting, and absorptive colonic cell lineages, respectively. The cells also possess some mesenchymal features, including vimentin and α- smooth muscle actin expression when proliferating actively. Finally, immortalized cells express various stem cell markers including LGR5, BMI1, CD29, and CD44. When cells are seeded at low density in Matrigel® in the absence of a mesenchymal feeder layer individual cells are able to divide and form self-organizing, cyst-like structures with a subset of cells exhibiting either mucin-2 or polarized villin staining. These findings demonstrate that the immortalized human colonic cells are capable of self renewal and multilineage differentiation. These cells should serve as valuable cellular reagents for studying normal colon stem cell biology, differentiation, and disease development.
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| Overall design |
Microarrays were performed on colonic epithelial cells from the two patients. These include:1. Log-growth phase early passage non-immortalized cells (controls). 2. Log-growth phase immortalized cells. 3. Immortalized cells in reduced serum (0.1% ) and reduced epithelial growth factor (EGF) (slow-growing cells). Immortalized cells in reduced serum (0.1% ), reduced EGF, and treated with GSK-3 beta inhibitor 6-bromoindirubin-3-oxime (BIO), (growth-arrested cells). duplicates were submitted for all four samples from each patient.
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| Contributor(s) |
Roig AI, Eskiocak U, Hight SK, Kim SB, Delgado O, Souza RF, Spechler SJ, Wright WE, Shay JW |
| Citation(s) |
19962984 |
| Submission date |
Oct 06, 2009 |
| Last update date |
Feb 18, 2019 |
| Contact name |
Andres I Roig |
| E-mail(s) |
andres.roig@utsouthwestern.edu
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| Phone |
214-648-6318
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| Fax |
214-648-8694
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| Organization name |
UT Southwestern
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| Department |
Cell Biology
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| Lab |
Shay/Wright lab
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| Street address |
5323 Harry Hines Blvd
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| City |
Dallas |
| State/province |
TX |
| ZIP/Postal code |
75390 |
| Country |
USA |
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| Platforms (1) |
| GPL6884 |
Illumina HumanWG-6 v3.0 expression beadchip |
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| Samples (16)
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| GSM459376 |
non-immortalized cells (2b HCEC1CT) |
| GSM459377 |
growth-arrested cells (3a HCE1CT+BIO) |
| GSM459378 |
growth-arrested cells (3b HCE1CT+BIO) |
| GSM459379 |
slow-growing cells (4a HCE1CT-BIO) |
| GSM459380 |
slow-growing cells (4b HCE1CT-BIO) |
| GSM459381 |
immortalized cells (5a HCEC2) |
| GSM459382 |
immortalized cells (5b HCEC2) |
| GSM459383 |
non-immortalized cells (6a HCEC2CT) |
| GSM459384 |
non-immortalized cells (6b HCEC2CT) |
| GSM459385 |
growth-arrested cells (7a HCE2CT+BIO) |
| GSM459386 |
growth-arrested cells (7b HCE2CT+BIO) |
| GSM459387 |
slow-growing cells (8a HCE2CT-BIO) |
| GSM459388 |
slow-growing cells (8b HCE2CT-BIO) |
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| Relations |
| BioProject |
PRJNA118233 |
| Supplementary file |
Size |
Download |
File type/resource |
| GSE18433_RAW.tar |
6.3 Mb |
(http)(custom) |
TAR |
| GSE18433_control_probes.txt.gz |
89.5 Kb |
(ftp)(http) |
TXT |
| GSE18433_non-normalized.txt.gz |
5.2 Mb |
(ftp)(http) |
TXT |
| Processed data included within Sample table |
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