1. Int J Oncol. 1998 Feb;12(2):257-63.

Establishment and characterization of immortalized human cell lines from
prostatic carcinoma and benign prostatic hyperplasia.

Iype PT, Iype LE, Verma M, Kaighn ME.

Biological Research Faculty and Facility, Inc., 10075-20 Tyler Place, Ijamsville,
Maryland 21754, USA.

New human prostate cell lines were developed from prostatic carcinoma (BRF-41T)
and BPH (BRF-55T). Primary cultures were initiated from cellular outgrowths of
explanted tissues. A serum-free medium, BRFF-HPC1, was developed for growing
human prostatic cancer cells. Cell strains were immortalized with pRSV-T plasmid 
to generate permanent cell lines that exhibited an epithelial morphology. Both
cell lines expressed the epithelial cell markers, cytokeratins 8 and 18 as well
as the prostatic marker, PSA, and the androgen receptor gene. They possess the
H-ras, K-ras, and p53 genes. We hope that these new human prostatic cell lines
will be useful as in vitro models for cancer research.

PMID: 9458347  [PubMed - indexed for MEDLINE]


2. Cytotechnology. 1998;26(3):207-18.

Development and characterization of new immortalized human breast cell lines

Iype LE, Michael M, Verma M, Iype PT.

Biological Research Faculty and Facility, Inc., Ijamsville, Maryland 21754, USA.

New human breast cell lines were developed from metastatic breast cancer tissues 
and normal breast tissues. Primary cultures were initiated from cellular
outgrowths of explanted tissues or from mechanically isolated cells in two
serum-free media. Cell cultures derived from both cancer and normal tissues were 
immortalized with pRSV-T plasmid to generate permanent breast cell lines that
exhibited an epithelial morphology. Cell lines generated in this study were
characterized with respect to morphology, growth rate, karyotype, presence of
specific genes, and the expression of epithelial and breast markers. The cell
lines expressed the epithelial cell markers, cytokeratins 8 and 18, and retained 
the capacity to produce human milk fat globulin. They also express the BRCA-1,
erbB2, and EGF receptor genes and possess the H-ras, K-ras, and p53 genes.
Preliminary data showed that one of the new cancer cell lines was highly
sensitive to the cytotoxic action of taxol. It is envisioned that the new breast 
cell lines will be useful as targets for identification of therapeutic agents
against breast cancer and as models for carcinogenesis studies.

PMCID: PMC3449551
PMID: 9618944  [PubMed - as supplied by publisher]


3. In Vitro Cell Dev Biol Anim. 1995 Jul-Aug;31(7):516-27.

Establishment of immortalized alveolar type II epithelial cell lines from adult
rats.

Driscoll KE, Carter JM, Iype PT, Kumari HL, Crosby LL, Aardema MJ, Isfort RJ,
Cody D, Chestnut MH, Burns JL, et al.

Procter and Gamble Company, Miami Valley Laboratories, Cincinnati, Ohio
45253-8707, USA.

We developed methodology to isolate and culture rat alveolar Type II cells under 
conditions that preserved their proliferative capacity, and applied lipofection
to introduce an immortalizing gene into the cells. Briefly, the alveolar Type II 
cells were isolated from male F344 rats using airway perfusion with a pronase
solution followed by incubation for 30 min at 37 degrees C. Cells obtained by
pronase digestion were predominantly epithelial in morphology and were positive
for Papanicolaou and alkaline phosphatase staining. These cells could be
maintained on an extracellular matrix of fibronectin and Type IV collagen in a
low serum, insulin-supplemented Ham's F12 growth medium for four to five
passages. Rat alveolar epithelial cells obtained by this method were transformed 
with the SV40-T antigen gene and two immortalized cell lines (RLE-6T and RLE-6TN)
were obtained. The RLE-6T line exhibits positive nuclear immunostaining for the
SV40-T antigen and the RLE-6TN line does not. PCR analysis of genomic DNA from
the RLE-6T and RLE-6TN cells demonstrated the T-antigen gene was present only in 
the RLE-6T line indicating the RLE-6TN line is likely derived from a spontaneous 
transformant. After more than 50 population doublings, the RLE-6T cells stained
positive for cytokeratin, possessed alkaline phosphatase activity, and contained 
lipid-containing inclusion bodies (phosphine 3R staining); all characteristics of
alveolar Type II cells. The RLE-6TN cells exhibited similar characteristics
except they did not express alkaline phosphatase activity. Early passage RLE-6T
and 6TN cells showed a near diploid chromosome number.(ABSTRACT TRUNCATED AT 250 
WORDS)

PMID: 8528500  [PubMed - indexed for MEDLINE]