cytokeratin 8 [46793]

CSF1PO: 13

D13S317: 8,14

D16S539: 9,11

D5S818: 12,15

D7S820: 10,11

THO1: 8,9.3

TPOX: 8,11

vWA: 14,18

ES-D, 2

G6PD, B

GLO-I, 1-2

Me-2, 0

PGM1, 2

PGM3, 1

RWPE-2 cells were derived from RWPE-1 cells (ATCC CRL-11609) by transformation with Ki-ras using the Kirsten murine sarcoma virus (Ki-MuSV) [PubMed: 9214605]. Epithelial cells from the peripheral zone of a histologically normal adult human prostate were transfected with a plasmid carrying one copy of the human papilloma virus 18 (HPV-18) genome to establish the RWPE-1 cell line (ATCC CRL-11609). RWPE-1 cells neither grow in agar nor form tumors when injected into nude mice with or without Matrigel. However, RWPE-2 cells form small colonies in soft agar and form tumors when injected into nude mice [PubMed: 9214605]. Further, a family of tumorigenic cell lines, that mimics multiple steps in prostate cancer progression, was also derived from RWPE-1 cells by exposure to N-methyl-N-nitrsourea (MNU). See the WPE1-NA22 (ATCCCRL-2849), WPE1-NB14 (ATCC CRL-2850), WPE1-NB11 (ATCC CRL-2851) and WPE1-NB26 (ATCC CRL-2852) cell ines.

The depositor reports that the RWPE-1 cell line (ATCC CRL-11609) was screened, and found negative for, Hepatitis B virus, Hepatitis C virus and Human immunodeficiency virus.

• 0.05 mg/ml BPE - provided with the K-SFM kit

• 5 ng/ml EGF - provided with the K-SFM kit. NOTE: Do not filter complete medium.
  Atmosphere: air, 95%; carbon dioxide (CO2), 5%
  Temperature: 37.0°C

1. Remove and discard culture medium.

2. Briefly rinse the cell layer with Ca++/Mg++ free Dulbecco's phosphate-buffered saline (D-PBS).

3. Add 2.0 to 3.0 ml (to a T-25 flask) or 3.0 to 4.0 ml (to a T-75 flask) of 0.05% Trypsin - 0.53mM EDTA solution, diluted 1:1 with D-PBS, and place flask in a 37C incubator for 5 to 8 minutes. Observe cells under an inverted microscope until cell layer is dispersed (usually within 5 to 10 minutes).

   Note: To avoid clumping do not agitate the cells by hitting or shaking the flask while waiting for the cells to detach.

4. Add 6.0 to 8.0 ml of 0.1% Soybean Trypsin Inhibitor (or 2% fetal bovine serum in D-PBS), as appropriate, and aspirate cells by gently pipetting.

5. Transfer cell suspension to centrifuge tube and spin at approximay 125 x g for 5 to 7 minutes.

6. Discard supernatant and resuspend cells in fresh serum-free growth medium. Add appropriate aliquots of cell suspension to new culture vessels. An inoculum of 2 X 10(4) to 4 X 10(4) viable cells/sq. cm is recommended.

7. Incubate cultures at 37C. We recommend that you maintain cultures at a cell concentration between 4 X 10(4) and 8 X 10(4) cells/sq. cm.

purified DNA:ATCC CRL-11610D

derived from same individual:ATCC CRL-2849

derived from same individual:ATCC CRL-2850

derived from same individual:ATCC CRL-2852

derived from same individual:ATCC CRL-2851

derivative:ATCC CRL-2853

derived from same individual:ATCC CRL-2854