See
“An Overview” for more information on the
derivation of these cell types.
Live
cells for distribution are sent frozen (in dry ice) in ampoules containing 5 x
10e5 or 1 x 10e6 cells. Under special circumstances, we can arrange to send
growing cells in flasks. For some cell types, RNA is also available upon
specific request.
See
"REVIEW
section VIII." for more information on cell shipments.
Note: I’ve been developing this system for over 30 years; it’s complicated!
Since I don’t expect others to know which of the many cell types generated would be best
for their studies, I recommend that you consult with me first about what available cells
may be most appropriate.
1. Pre-stasis (Chart
1, Panel C)
Reduction
mammoplasty derived HMEC grown in a serum-containing medium, M87A or M85 (Stampfer
et al. 1980; Stampfer 1982; Garbe et al. 2009; Garbe et al. 2012; LaBarge et al. 2013; Stampfer et al. 2013; Sputova et al. 2013; Pelissier et al. 2014; Garbe et al. 2014)
We
have large batches of pre-stasis HMEC to distribute at passages 4-8; most
growth ceases around passages 13-16. Lower passage amps may be sent to allow growth of stocks in the
recipient’s lab. Pre-stasis HMEC contain a range of phenotypes, particularly at
lower passages; cells with luminal or progenitor markers make up ~5-30% of the
population. Pre-stasis HMEC are
genomically stable, even at stasis. I
most commonly distribute cells from specimens 184, 48R, and 240L (ages
16-21). More limited quantities of HMEC
derived from older woman are also available.
2. Post-stasis: BaP (previously called Extended Life) (p16 silenced/mutated)
(Chart 1, Panel A)
Reduction
mammoplasty derived HMEC from specimen 184 that were exposed to the chemical
carcinogen benzo(a)pyrene in primary culture.
(Stampfer
& Bartley 1985, 1988; Brenner et al., 1998; Novak et al. 2009; Severson et al. 2014)
Very limited quantities of the BaP cultures are available. These include: 184Aa (p16 mutated), the precursor of 184A1 and other 184A- lines; 184Be (p16 silenced), the precursor of 184BE1 and 184BE2 lines, and 184Ce, the precursor of the184CeMY line. Talk with me directly about these cells. More 184Aa is available than any other BaP type. These cells were exposed to BaP and so harbor many point mutations.
3. Post-stasis: Post-selection (p16 silenced) (Chart 1, Panel B)
HMEC grown in a high stress-inducing, serum-free medium, that emerge from p16(+) populations at stasis following silencing of the p16 promoter.
(Hammond et al. 1984; Romanov et al. 2001; Garbe et al. 2007, 2009, 2014; Li et al. 2007; Brenner et al., 1998; Novak et al. 2009)
We
have large batches of post-selection HMEC frozen at around passages 7-10 that
we distribute. Depending upon the individual, these cells cease active growth
around passages 14-25 (about 3 PD per passage).
These cells are routinely available from women of different ages. I most commonly distribute cells from
specimens 184, 48R, 239, 240L and 161.
These cells are not normal. They have overcome the stasis barrier and show many differences from normal pre-stasis HMEC (e.g., in gene expression and promoter methylation), some of which are also seen in breast cancer cells. They express a generally basal phenotype, although some luminal marker may also be present at higher passages (e.g., mucins and keratin 18), and they exhibit metaplastic properties (Sauder et al. BMC Biology 2014). They become genomically unstable as they approach the telomere dysfunction senescence barrier. Post-selection HMEC have also been referred to as vHMEC, and suggested to give rise to metaplastic cancer (Keller et al. PNAS 2012). This is the cell type sold commercially as “normal primaries” (e.g., Lonza CC-2551 and Life Technologies A10565) although these significantly aberrant cells are neither normal nor primary.
4. Post-stasis: GSE22 (p53 expression inhibited by GSE22)
(Chart 1, Panel A)
Pre-stasis
HMEC grown in MM that were transduced with GSE22 and show clonal escape from stasis.
In
2 separate experiments pre-stasis 184 HMEC were transduced with a genetic
suppressor element (GSE22) that produces a peptide that interferes with p53
function. In both cases there were a
few clonal outgrowths when the vast majority of the cells ceased growth at
stasis. These cells have not been
well-characterized. They express low but
detectable p16. These cells are not routinely distributed.
5. Post-stasis: p16sh (p16 expression inhibited by shRNA
to p16) (Chart 1, Panel C)
Pre-stasis HMEC grown in a low stress-inducing medium that were exposed to shRNA to p16 and show uniform bypass of stasis.
Pre-stasis 184, 240L, 122L, and 805P HMEC were transduced with shRNA to p16 at early passages (184F batch in M85 medium; the others in M87A+X medium). These cells have not been well-characterized. They express increased telomerase (TRAP) activity compared to the parental pre-stasis HMEC. These cells are not routinely distributed but can be made available.
B. FINITE LIFESPAN HUMAN MAMMARY FIBROBLASTS (HMF) (Stampfer et al. 1981; Bartley et al., 1982; Romanov et al. 2001; Vrba et al. 2010, 2011; Novak et al. 2012)
We
have available for distribution stocks of fibroblast cells from several
reduction mammoplasty specimens for which HMEC are available, e.g., specimens
184, 48, 240, and 161. These cells are grown in a serum-containing medium. In theory, fibroblast stocks can be obtained
from any of our specimens, including the mastectomy derived tissues, but we
have not grown up stocks to distribute from more than a few. Frozen cells are available around passages
5-8, and they senesce around passages 12-20 (2-3 PD per passage) depending upon
the individual. These cells grow slower
than the HMEC, and we do not generally have as large stocks available.
We
have very limited quantities of
frozen organoids, and are therefore very reluctant to distribute any of this
material, but will make exceptions for specific studies. More primary tissue is
available from reduction mammoplasties than mastectomies. Talk with me directly
about these cells.
D.
IMMORTALLY TRANSFORMED CELL LINES
(Stampfer & Bartley 1985, 1988; Walen & Stampfer 1989;
Lehman et al. 1993; Brenner et al. 1998; Stampfer et al. 1997, 2001, 2003;
Garbe et al. 1999, 2007, 2014; Nonet et al. 2001; Olsen et al. 2002; Chin et al.
2004; Li et al. 2007; Novak et al. 2009; Pelissier et al. 2014; Severson et al. 2014)
1. IMMORTAL LINES DERIVED FROM CELLS GROWN IN MM, FOLLOWING EXPOSURE TO ONCOGENIC AGENTS (Chart 1, Panel A)
1A) Lines derived from cells exposed to benzo(a)pyrene (BaP); i.e. from the BaP post-stasis cultures:
184A1 (Stampfer & Bartley
1985)
Unlimited
quantities of later passages of this immortal cell line are available for
distribution. Very limited quantities of
early passage newly immortal (pre-conversion) cells are available upon specific
request. Clonal isolates are also
available. These cells are wild type for
p53 and RB, and are not anchorage independent or tumorigenic. They have the most stable karyotype of our BaP-exposed immortally transformed lines. This line is derived from the BaP post-stasis precursor 184Aa (p16 mutated).
184A1-RF (Olsen et al. 2002)
184A1was transduced with an activated Raf-1 construct under the control of inducible ER. These cells now have AIG and reduced EGF requirements for growth.
184A1-GSE22 (Stampfer et al. 2003)
184A1was
transduced with GSE22. When the GSE22
is introduced pre-conversion (passage 12) conversion proceeds rapidly. When the
GSE22 is introduced into fully immortal 184A1 it can provide a matched p53(-)
culture to the p53(+) 184A1. These cells are not routinely distributed but can
be made available.
184A1-hTERT(12p) (Stampfer et al. 2001; Olsen et al 2002)
Early
passage pre-conversion (passage 12) 184A1 was transduced with hTERT, resulting
in immediate uniform immortalization without undergoing conversion. These cells are not routinely distributed.
184A1-hTERT(22p) (Stampfer et al. 2001)
184A1
already in the process of conversion (passage 22) was transduced with
hTERT. These cells proceeded through
conversion to become fully immortal. These cells are not routinely distributed.
184A1- E6; -E7; -T; -E1A (Garbe et
al. 1999)
Early passage pre-conversion (passage 12) 184A1 was transduced with either the HPV16 -E6, -E7, SV40T, or E1A genes. These viral oncogenes produce numerous effects that may accelerate or alter the conversion process. These cells are not routinely distributed.
184AA4 (Stampfer et al. 2003)
Unlimited
quantities of later passages of this immortal line are available for
distribution. Early passage cells are
not available. These cells are wild type for p53, and RB, and are not anchorage
independent or tumorigenic. They have
many karyotypic abnormalities. This line is derived from the BaP post-stasis precursor 184Aa.
184AA2 (Stampfer et al. 2003)
Unlimited
quantities of this p53(-/-) immortal line are available for distribution. These
cells are wild type for RB, and have AIG.
The karyotype is unstable. 184AA2 is derived from the same BaP post-stasis precursor population (184Aa) as 184A1 and 184AA4. 184Aa was transduced with retroviral vectors that gave insertional mutagenesis at the p53 locus. We presume this line acquired addition errors at crisis.
184AA3 (Stampfer et al. 2003)
Unlimited quantities of this p53(-/-) immortal line are available for distribution. These cells are wild type for RB, and have AIG by passage 50. The karyotype is unstable. 184AA3 is derived from the same BaP post-stasis precursor population (184Aa) as 184A1 and 184AA4. 184Aa was transduced with retroviral vectors that gave insertional mutagenesis at the p53 locus. We presume this line acquired addition errors at crisis.
184AA5-7
These are immortal lines that arose in finite lifespan BaP post-stasis 184Aa populations transduced with control retroviruses. They have not been characterized, but their early rapid growth suggests they are p53(-) like 184AA2 and 184AA3, and are a consequence of insertional mutagenesis. These cells are not routinely distributed.
184AA8
This is a new line of indefinite lifespan that emerged from BaP post-stasis 184Aa. It is not yet well-characterized; it’s gradual conversion process is consistent with a p53(+) status. It is not yet available for distribution.
184AaGS1-2
Finite lifespan BaP post-stasis 184Aa was transduced with GSE22 and produced two immortally transformed cultures. Immortalization is clonal, and each isolate is likely to be somewhat different. We presume these lines have acquired addition errors at crisis. These lines have not been well-characterized and are not routinely distributed.
184AaMY1-5 (Garbe et al. 2014)
Finite lifespan BaP post-stasis 184Aa was transduced with c-myc on multiple occasions; each time there has been efficient rapid immortalization of the population. These lines have not been well-characterized. They are not routinely distributed but can be made available.
184AaZN1-3 (Nonet et al. 2001)
Finite lifespan BaP post-stasis 184Aa was transduced with the breast cancer-associated oncogene ZNF217 on multiple occasions. Immortalization is clonal, and each isolate is likely to be somewhat different. We presume these lines have acquired additional errors during the period of agonescence. The lines examined are p53(+). They are not routinely distributed but can be made available.
184AaE6 (Garbe et al. 1999)
Transduction of the HPV16 E6 gene into BaP post-stasis 184Aa produces common immortalization following crisis. These cells are not routinely distributed but can be made available.
184B5 (Stampfer & Bartley
1985)
Unlimited quantities of later passages of this immortal line are available for distribution. More limited quantities of early passage newly immortal (pre- and mid- conversion) cells are available upon specific request. Clonal isolates are also available. These cells are wild type for p53, and RB, and are not anchorage independent or tumorigenic. They have a low level of karyotypic instability. We previously thought that this line was derived from BaP post-stasis 184Be, however recent studies (Severson et al. and in prep) indicate that it arose from a minor (~10%) population in the 184Be culture, which we have now named 184Bd. Unfortunately, pure cultures of 184Bd are not available.
184B5-erbB2
184B5
was transfected with the breast cancer associated oncogene erbB2; these cells
exhibit AIG.
184B5ME was isolated from anchorage
independent colonies of 184B5-erbB2 grown in methocel. They have not been well-characterized. Unlimited quantities are available.
184BE1 (Severson et al. 2014)
This is a new line of indefinite lifespan that emerged from BaP post-stasis 184Be. It is not yet well-characterized; it’s gradual conversion process is consistent with a p53(+) status. This line is not routinely distributed but can be made available.
184BE2
This is a new line of indefinite lifespan that emerged from BaP post-stasis 184Be. It is not yet well-characterized; it’s gradual conversion process is consistent with a p53(+) status. It is not yet available for distribution.
184BEMY1 (Garbe et al. 2014)
Finite lifespan BaP post-stasis 184Be was transduced with c-myc, producing efficient rapid immortalization of the population. This line has not been well-characterized and is not yet available for distribution.
184CEMY1 (Garbe et al. 2014)
Finite lifespan BaP post-stasis 184Ce was transduced with c-myc, producing efficient rapid immortalization of the population. This line has not been well-characterized and is not yet available for distribution. We have not derived any “spontaneous” lines from 184Ce (culture of 184Ce has been more limited than 184Aa or 184Be).
1B) Line derived from cells exposed to GSE22
184FGS1 (Garbe et al. 2007)
An immortal line emerged
from one of the two post-stasis 184F-GSE22 cultures. These cells have not been well-characterized.
This line is not routinely distributed but can be made available.
1C) Line derived from cells exposed to hTERT
184FTERT (Stampfer et al. 2001)
Pre-stasis 184F grown in MM
were transduced with hTERT at passage 3. A clonal population maintained growth
after the
vast majority of the cells ceased growth at stasis. The population
gradually lost expression of p16 while gaining resistance to TGFß growth
inhibition. These cells have not been
well-characterized. This line is not routinely distributed but can be
made available.
2. IMMORTAL LINES DERIVED FROM POST-SELECTION POST-STASIS HMEC GROWN IN SERUM-FREE MCDB170
(Chart
1, Panel B)
2A) Lines derived from post-selection HMEC transduced with c-myc and/or ZNF217 and/or GSE22
184ZN4-7 (Nonet et al. 2001; Chin et
al. 2004)
Finite
lifespan post-selection 184 was transduced with ZNF217 on multiple
occasions. In 7 independent experiments 4 clonal immortal lines
emerged;
each isolate is likely to be somewhat different. We presume these lines acquired additional
errors during the period of agonescence. The lines examined are p53(+).These lines are not routinely distributed but can be
made available.
Finite
lifespan post-selection 184 was transduced with ZNF217, followed by c-myc, on
multiple occasions. In 5 independent experiments 4 clonal immortal lines
emerged. Three of these lines emerged
soon after the exposure to c-myc, prior to agonescence; Southern analysis
indicated they are clonal. The 2 lines
examined by array CGH show no alterations in copy numbers. We presume no additional genomic mutations
were required for immortalization. These
lines are
not routinely distributed but can be made available on a collaborative basis
upon request.
184ZNMY3-N
We
have transduced 184ZNMY2 and 184ZNMY3 with oncogenic erbB2 (neu). These lines then acquired AIG. CGH analysis
shows no copy number alterations.
184ZNGS1
Finite lifespan post-selection 184 was transduced with ZNF217, followed byGSE22, in two experiments. In 4 independent exposures one immortal line emerged soon after the exposure to GSE22, prior to agonescence. These cells have not been well-characterized. This line is not routinely distributed but can be made available.
2B) Lines derived from post-selection HMEC transduced with hTERT
184BTERT; 48RTERT; 161HTERT
Transduction
of hTERT into post-selection p16(-) HMEC produces efficient
immortalization bypassing the genomic instability of agonescence,
and conversion. Unlimited quantities of 184BTERT are available for
distribution.
2C) Line derived from post-selection HMEC transduced with HPV-E6
184-E6
Transduction of the HPV16 E6 gene into post-selection p16(-) HMEC produces common immortalization following crisis, bypassing conversion. These cells are not routinely distributed but can be made available.
3. IMMORTAL LINES DERIVED FROM CELLS GROWN IN M85/M87/M87A ± OXYTOCIN (X) (Chart 1, Panel C)
These lines can be made available on a collaborative basis upon request.
3A) Lines derived following transduction with c-myc and/or shRNA to p16 (Garbe et al. 2014; Lee et al. submitted)
184FMY2, 184DMY3, 240LMY, 122LMY (clonal)
Pre-stasis 184F growing in M85, and pre-stasis 184D, 240LB, and 122L growing in M87A+X, were transduced with c-myc. There was clonal escape from stasis, yielding immortal lines.
184Fp16s, 184Dp16s, 240Lp16s, 805Pp16s (clonal)
Pre-stasis 184F growing in M85, and pre-stasis 184D, 240LB and 805P growing in M87A+X, were transduced with shRNA to p16, producing p16sh post-stasis populations. Clonal immortal lines emerged around the period of agonescence.
184Fp16sMY, 184Dp16sMY, 240Lp16sMY, 122Lp16sMY, 805Pp16sMY (non-clonal)
Pre-stasis 184F growing in M85, and pre-stasis 184D, 240LB, 122L, and 805P growing in M87A+X, were transduced with shRNA to p16, then c-myc, producing increased telomerase activity and uniform immortalization.
3B) Lines derived following transduction with hTERT (Garbe et al. 2014)
184DTERT
Pre-stasis
184D growing in M87A+X were transduced with hTERT and showed non-clonal immortalization.
Selected
Publications:
Stampfer, M.R., Hallowes, R. and Hackett, A.J., Growth of Normal Human Mammary Epithelial Cells in Culture. In Vitro 16:415-425, 1980. PMID: 6993343
Stampfer, M.R., Bartholomew, J.C., Smith, H.S., and Bartley, J., Metabolism of Benzo(a)Pyrene by Human Mammary Epithelial Cells: Toxicity and DNA Adduct Formation. Proc. Natl. Acad. Sci. (USA) 78:6251-6255, 1981. PMID: 6273860
Bartley, J.C., Bartholomew, J.C. and Stampfer, M.R., Metabolism of Benzo(a)pyrene in Human Mammary Epithelial and Fibroblast Cells: Metabolite Pattern and DNA Adduct Formation. J. Cell. Biochem. 18:135-148, 1982. PMID: 6279686
Stampfer, M.R., Cholera Toxin Stimulation of Human Mammary Epithelial Cells in Culture. In Vitro 18:531-537, 1982. PMID: 6288550
Hammond, S.L., Ham, R.G., and Stampfer, M.R., Serum-free Growth of Human Mammary Epithelial Cells: Rapid Clonal Growth in Defined Medium and Extended Serial Passage with Pituitary Extract. Proc. Natl. Acad. Sci. (USA) 81:5435-5439, 1984. PMID: 6591199
Stampfer, M.R., and Bartley, J.C., Induction of Transformation and Continuous Cell Lines from Normal Human Mammary Epithelial Cells after Exposure to Benzo(a)pyrene. Proc. Natl. Acad. Sci. (USA) 82:2394-2398, 1985. PMID: 3857588
Stampfer, M.R., and Bartley, J.C., Human Mammary Epithelial Cells in Culture: Differentiation and Transformation. Cancer Treat Res 40: 1-24, 1988. PMID: 2908646
Walen, K.H., and Stampfer, M.R., Chromosome Analyses of Human Mammary Epithelial Cells (HMEC) at Stages of Chemically-induced Transformation Progression to Immortality, Cancer Genet. Cytogen. 37:249-261, 1989. PMID: 2702624
Lehman T, Modali R, Boukamp P, Stanek, J., Bennett, W.P., Welsh, J.A., Metcalf, R.A., Stampfer, M.R., Fusenig, N., Rogan, E.M., Reddel, R., and Harris, C.C. p53 mutations in human immortalized epithelial cell lines. Carcinogenesis 14, 833-839, 1993. PMID: 8504475
Stampfer, MR, Bodnar, A, Garbe, J, Wong, M, Pan, A, Villeponteau, B, Yaswen, P, Gradual phenotypic conversion associated with immortalization of cultured human mammary epithelial cells, Mol Biol Cell 8:2391-2405, 1997. PMID: 9398663
Brenner, AJ, Stampfer, MR, Aldaz, M, Increased p16 expression with first senescence arrest in human mammary epithelial cells and extended growth capacity with inactivation, Oncogene 17:199-205, 1998. PMID: 9674704
Garbe, J, Wong, M, Wigington,
D, Yaswen, P, Stampfer, MR, Viral oncogenes accelerate conversion to
immortality of cultured conditionally immortal human mammary epithelial cells,
Oncogene 18:2169-2180, 1999. PMID: 10327063
Romanov, SR, Kozakiewicz, K, Holst, CR, Stampfer, MR,
Haupt, LM, Tlsty, TD, Normal human mammary epithelial cells spontaneously
escape senescence and acquire genomic changes, Nature 409:633-637, 2001. PMID:
11214324
Nonet, GH, Stampfer, M, Chin, K,
Gray, JW, Collins, CC, Yaswen, P, The ZNF217 Gene amplified in breast cancers
promotes immortalization of human mammary epithelial cells, Cancer Res. 61:
1250-1254, 2001. PMID: 11245413
Stampfer, MR, Garbe, J, Levine, G,
Lichtsteiner, S, Vasserot, AP, Yaswen, P, hTERT expression can induce
resistance to TGFß growth inhibition in p16INK4A(-)
human mammary epithelial cells, Proc Natl Acad Sci (USA) 98:4498-4503, 2001. PMID:
11287649
Olsen CL, Gardie, B, Yaswen, P, Stampfer, MR,
Raf-1-induced growth arrest in human mammary epithelial cells is
p16-independent and is overcome in immortal cells during conversion, Oncogene
21:6328-6339, 2002. PMID: 12214273
Stampfer, M, Garbe, J, Nijjar, T, Wigington, D, Swisshelm, K, Yaswen, P, Loss of p53 function accelerates acquisition of telomerase activity in indefinite lifespan human mammary epithelial cell lines, Oncogene 22:5238-5251, 2003. PMID: 12917625
Chin, K, Ortiz de Solorzano, C, Knowles, D, Jones, A, Chou,W, Rodriguez, E, Kuo, W-L, Ljung, B-M, Chew, K, Krig, S, Garbe, J, Stampfer, M, Yaswen, P, Gray, JW, Lockett, SJ. In Situ Analysis of Genome Instability in Breast Cancer. Nature Genetics:36, 984-988 2004. PMID: 15300252
Li Y, Pan J, Li J-L, Lee J-H, Tunkey C, Saraf K, Garbe J, Jelinsky S, Stampfer MR, Haney, SA, Transcriptional Changes Associated with Breast Cancer Occur as Normal Human Mammary Epithelial Cells Overcome Senescence Barriers and Become Immortalized. Mol Can 6:7, 2007. PMID: 17233903
Garbe, J, Holst, CR, Bassett, E, Tlsty, T, Stampfer, MR, Inactivation of p53 Function in Cultured Human Mammary Epithelial Cells Turns the Telomere-Length Dependent Senescence Barrier from Agonescence into Crisis. Cell Cycle 6:1927-1936, 2007. PMID: 17671422
Novak, P, Jensen, TJ, Garbe, JC, Stampfer, MR, Futscher, BW, Step-wise DNA methylation changes are linked to escape from defined proliferation barriers and mammary epithelial cell immortalization, Cancer Res 69:5251-58, 2009. PMID: 18922938
Garbe JC, Bhattacharya S, Merchant B, Bassett E, Swisshelm K, Feiler HS, Wyrobek AJ, Stampfer MR, Molecular distinctions between the stasis and telomere attrition senescence barriers demonstrated by long-term culture of normal human mammary epithelial cells. Cancer Res 69:7557-7568, 2009. PMID: 19773443
Vrba, L, Jensen, TJ, Garbe, JC, Heimark, RL, Cress, AE, Dickinson, S, Stampfer, MR, Futscher, BW, DNA Methylation Control of Normal Cell-Type Specific Expression of miR-200c, PLoS ONE 5(1): e8697, 2010. PMID: 20084174
Vrba L, Garbe JC, Stampfer MR, Futscher BW. Epigenetic regulation of normal human mammary cell type specific miRNAs. Genom Res 21:2026-2037, 2011. PMID: 21873453
Garbe JC, Pepin F, Pelissier F, Sputova K, Fridriksdottir A, Guo DE, Villadsen R, Park M, Petersen OW, Barowsky A, Stampfer MR, LaBarge MA. Accumulation of multipotent progenitors with a basal differentiation bias during aging of human mammary epithelia. Cancer Res 72:3687-701, 2012. PMID: 22552289
Novak P, Stampfer MR, Munoz-Rodriguez JL, Garbe JC, Ehrich M , Futscher BW Jensen TJ.
Cell-Type Specific DNA Methylation Patterns Define Human Breast Cellular Identity. PLoS One 8(2): e53776, 2012. e52299. PMID: 23284978
LaBarge MA, Garbe JC, Stampfer MR. Processing of human reduction mammoplasty and mastectomy tissues for cell culture. J Vis Exp 71: e50011 2013. PMID: 23328888
Vrba L, Muñoz-Rodríguez JL, Stampfer MR, Futscher BW. miRNA Gene Promoters Are Frequent Targets of Aberrant DNA Methylation in Human Breast Cancer. PLoS One 8(1): e54398, 2013. PMID 23342147
Stampfer MR, LaBarge MA, Garbe JC. An Integrated Human Mammary Epithelial Cell Culture System for Studying Carcinogenesis and Aging, in: Cell and Molecular Biology of Breast Cancer, ed. H. Schatten, Springer, NY pp323-361, 2013.
Sputova, K, Garbe, JC, Pelissier, FA, Chang, E, Stampfer, MR, Labarge, MA. Aging phenotypes in cultured normal human mammary epithelial cells are correlated with decreased telomerase activity independent of telomere length. Genome Integr 4:4 2013. PMID 23718190
Pelissier, FA, Garbe, JC, Ananthanarayanan, B, Miyano, M, Lin, C, Jokela, T, Kumar, S, Stampfer, MR, Lorens, JB, LaBarge, MA (2014). Age-related dysfunction in mechano-transduction impairs differentiation of human mammary epithelial progenitors. Cell Rep 7:1926-39. PMID 24910432
Garbe, JC, Vrba, L, Sputova, K, Fuchs, L, Novak, P, Jackson, MW, Chin, K, LaBarge, MA, Watts, GS, Futscher, BW, Stampfer, MR. Immortalization of normal human mammary epithelial cells in two steps by direct targeting of senescence barriers does not require gross genomic alterations. Cell Cycle, 13:3423-35, 2014. PMID 25485586.
Severson, PL, Vrba, L, Stampfer, MR, Futscher, BW. Exome-wide mutation profile in benzo[a]pyrene-derived post-stasis and immortal human mammary epithelial cells. Mut Res - Gen Tox Env Mut, 2014 in press. PMID 25435355
1. Pre-stasis (Chart
1, Panel C)
2. Post-stasis: BaP (formerly EL) (p16
silenced/mutated) (Chart 1, Panel A)
Reduction
mammoplasty derived HMEC from specimen 184 that were exposed to the chemical
carcinogen benzo(a)pyrene in primary culture.
3. Post-stasis: Post-selection (p16 silenced) (Chart 1, Panel B)
Reduction
mammoplasty, non-tumor mastectomy, or benign tumor derived HMEC, grown in a
serum-free medium, that emerge from populations at stasis following silencing
of the p16 promoter.
4. Post-stasis: GSE22 (p53 expression inhibited by GSE22)
(Chart 1, Panel A)
Pre-stasis
HMEC grown in MM that were exposed to GSE22.
5. Post-stasis: p16sh (p16 expression inhibited by shRNA
to p16) (Chart 1, Panel C)
Pre-stasis
HMEC grown in a serum-containing medium that were exposed to shRNA to p16 and
show uniform bypass of stasis.
B.
FINITE LIFESPAN HUMAN MAMMARY FIBROBLAST CELLS
(HMF)
D.
IMMORTALLY TRANSFORMED CELL LINES
1.
Immortal lines derived from cells grown in MM, following exposure to oncogenic
agents (Chart 1, Panel A)
1a) Lines derived from cells exposed to benzo(a)pyrene; i.e. from the BaP post-stasis cultures
184A1
184A1-RF
184A1-GSE22
184A1-hTERT(a)
184A1-hTERT(b)
184A1- E6; -E7; -T; -E1A
184AA4
184AA2
184AA3
184AA5-7
184AaGS1,2
184AaMY1-5
184AaZN1-3
184AaE6
184B5
184B5-erbB2
184B5ME
184BE1
184BE2
184BEMY1
184CEMY1
1b)
Line derived from cells exposed to GSE22
184FGS1
1c)
Line derived from cells exposed to hTERT
184FTERT
2.
Immortal lines derived from post-selection post-stasis HMEC grown in high stress serum-free MCDB170 (Chart
1, Panel B)
2a)
Lines derived from post-selection HMEC transduced with c-myc and/or ZNF217
184SMY1
184ZN4-7
184ZNMY3-N
184ZNGS1
2b)
Lines derived from post-selection HMEC transduced with hTERT
184BTERT; 48RTERT; 161HTERT
2c)
Line derived from post-selection HMEC transduced with HPV-E6
184-E6
3.
Immortal lines derived from cells grown in low stress low serum-containing
M85/M87/M87A media ± oxytocin (X) (Chart 1, Panel C)
3a)
Lines derived following transduction with c-myc and/or shRNA to p16
184FMY2
184DMY3
240LMY1
122LMY1
184Fp16s
184Dp16s
240Lp16s
805Pp16s
184Fp16sMY
184Dp16sMY
240Lp16sMY1
122Lp16sMY1
805Pp16sMY
3b)
Lines derived following transduction with hTERT
184DTERT