|
| Status |
Public on Aug 29, 2008 |
| Title |
Conjunctival_clone1 |
| Sample type |
RNA |
| |
|
| Source name |
Conjunctival keratinocyte clone from biopsy
|
| Organism |
Sus scrofa |
| Characteristics |
Passage V conjunctival keratinocyte clone
Generates goblet cells in culture
Expresses stem cell markers (p63, Zfp145, Bmi1), marker of corneal differentiation (K12) and of conjunctival differentiation (Muc5ac, K4 and K19)
|
| Growth protocol |
Biopsies of conjunctiva or cornea taken at distance (6mm) from the limbus were obtained from white domestic pigs (AP1 and AP6) soon after sacrifice. Biopsies were dissociated using 0.05 % trypsin in 0.01% EDTA and the keratinocytes cultivated on a feeder layer of irradiated 3T3-J2 cells in a 3:1 DMEM F12 medium supplemented as described (Oshima et al., Cell 2001). Single cells were isolated after the mass cultures with an elongated Pasteur pipet under an inverted microscope as described (Barrandon and Green Proc Natl Acad Sci USA, 1985) and cultivated as above.
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Total RNA was prepared using the RNeasy Mini Kit (QIAGEN Inc.)
|
| Label |
biotin
|
| Label protocol |
1 micro-gramme of total RNA were amplified and labeled using Affymetrix eukaryotic taget preparation (one-cycle amplification, #701025 Rev.2)
|
| |
|
| Hybridization protocol |
Affymetrix Porcine genome arrays were hybridized with 11 mg of labeled, amplified cRNA , washed, stained and scanned according to the protocol described in Affymetrix GeneChip® Expression Analysis Manual (Fluidics protocol EukGeWS2v5_450)
|
| Scan protocol |
Scanning was done on an Affymetrix GeneChip Scanner 7G
|
| Description |
CJ_1
|
| Data processing |
Normalized expression signals were calculated from Affymetrix CEL files using RMA (Irizarry et al., 2003).
|
| |
|
| Submission date |
Aug 28, 2008 |
| Last update date |
Aug 28, 2008 |
| Contact name |
Sylvain Pradervand |
| E-mail(s) |
Sylvain.Pradervand@unil.ch
|
| Phone |
+41 21 692 39 08
|
| Organization name |
UNI Lausanne
|
| Department |
CIG
|
| Lab |
DNA Array Facility
|
| Street address |
Genopode
|
| City |
Lausanne |
| ZIP/Postal code |
1015 |
| Country |
Switzerland |
| |
|
| Platform ID |
GPL3533 |
| Series (1) |
| GSE12604 |
Comparison of corneal and conjunctival keratinocyte clones |
|
