Vol.1, No.2, 21-28 (2011)
opyright © 2011 SciRes. Openly accessible at http://www.sc irp.org/journal/SCD/
Stem Cell Discovery
Establishment and molecular characterization of breast
cancer mesenchymal stem cell line derived from human
non-metastasis breast cancer tumor
Pravin Potdar, Sachin Chaugule
Department of Molecular Medicine and Biology, Jaslok Hospital and Research Centre, Mumbai, India;
Corresponding Author: ppotdar@jaslokhospital.net
Received April 29th, 2011; revised May 23rd, 2011; accepted May 31st, 2011.
Breast cancer remains a leading cause of mor-
bidity and mort ality in women mainly because o f
the propensity of primary breast tumors to me-
tastasize. It is composed of heterogeneous cell
populations with different biological properties.
Breast cancer-initiating c ells have been rec ently
identified in breast carcinoma as CD44+/CD24/low
cells, which display stem cell like properties. In
the present study, we have isolated breast can-
cer stem cells from non-met astasis tumor tissue,
which is presently at passage 18 and designated
as human Breast Cancer Mesenchymal Stem
Cells (hBCMSCs) line. These cells showed spin-
dle shaped morphology and formed mammos-
pheres as well as pluripotency clones indicating
their stem cell nature. Molecular marker study
confirmed mesenchymal nature as well as
pluripotency, plasticity and oncogenicity of
these cells. The hBCMSCs cell line may likely
contain a heterogeneous population of malig-
nant cells. Interestingly, we also found that
these cells exhibit BRCA 2 mutation, which was
found in Indian population. Overall, this study
revealed that hBCMSCs cell line may represent
a suitable in vitro model to study the mecha-
nism of breast cancer which further leads to an
identification of molecular targets for future
breast cancer targeted therapy.
Keywords: Breast Cancer Stem Cells; Breast
Cancer, BRCA2; Mesenchymal Stem Cells;
Molecular Markers
A fundamental problem in breast cancer research is to
identify the cell type capable of sustaining the growth of
the neoplastic clone. It is less clear from most of the
breast cancers studies that which cells within the tumor
clone possess tumor-initiating cell function for main-
taining tumor growth. The identification of potential
breast cancer stem cells is the most important task for
the study due to the characteristics of these stem cells
which are resistant to conventional cancer therapy [1].
Pardal et al. [2003] have shown that a small subset of
cells within a tumor, termed Cancer Stem Cells (CSCs)
or tumor-initiating cells which are responsible for tumor
initiation and tumor growth progression [2]. The concept
about tumor containing heterogeneous populations of
cells was demonstrated firstly by Lapidot and colleagues
in leukemia [3]. Fidler and Kripke observed striking
heterogeneity of dissociated tumor cells with respect to
the clonogenic ability to form metastasis [4]. The exis-
tence of “cancer stem cells” or “cancer initiating cells”
has been identified in a variety of solid tumors including
gliomas, medulloblastoma, lung cancer, prostate cancer,
colon cancer and breast cancer by various investigators
[5-11]. Recent studies have suggested the existence of
stem cells in the human mammary gland similar to those
in the rodent and also shown that non-adherent mam-
mospheres are highly enriched for cells with stem and
progenitor cell properties [12,13]. The isolation and
characterization of mammary cancer stem cells also has
important implications for understanding mammary car-
cinogenesis. Significant progress has already been made
in the identification of the stem cell in mammary gland
of the mouse, though relatively little work has been car-
ried out in humans [1]. It may help to design therapies
targeted to the unique properties of the tumor stem cell
to get selective killing. There is a need of comparing
Breast cancer initiating cells phenotypes with non-breast
cancer initiating cells phenotypes. The focus of future
studies involves cell signaling pathways, molecular and
cellular markers for normal and tumorigenic cells and
drug development.
P. Potdar et al. / Stem Cell Discovery 1 (2011) 21-28
Copyright © 2011 SciRes. Openl y accessible at http://www.scirp.org/journal/SCD/
Molecular marker study is one of the important stud-
ies to characterize and establish any cell type. Recent
studies on stem cell research have identified CD105,
CD13 and CD73 as a Mesenchymal Stem Cells (MSCs)
markers [14], CD34, CD45 as a Hematopoietic Stem
Cells (HSCs) markers [15,16], Oct4, Nanog, SOX2 as a
Pluripotency and self renewal capacity markers [17]. It
has been established that there is change in expression of
molecular markers like DAPK, COX2 and LIF in can-
cerous cell types [18-20]. CD44 is a receptor for halu-
ronic acid, which has been shown to play a role in tumor
migration and metastasis [21]. CD24 has recently been
described as a negative regulator of CXCR4, a chemo-
kine receptor important in facilitating breast cancer me-
tastasis [22]. An understanding of the biology of the cells
that drive tumorogenesis has the potential to lead to new
therapeutic approaches for breast cancer.
In present study, we have mainly focused on isolation,
characterization and propagation of in vitro breast cancer
stem cells from non-metastasis human breast tumor.
Moreover, our research further confined to the isolation
of breast cancer stem cells and characterizes these cells
for their mesenchymal or hematopoietic phenotypes as
well as pluripotency, oncogenic, cytokines and chemoki-
nes markers. As our laboratory has already established
Breast cancer mutation in BRCA2 gene in Indian popu-
lation [23]. We have decided to study this mutation in
vivo tumor tissue and in vitro isolated hBCMSCs cell
line to confirm the specificity of Breast cancer tumor
cells. Overall, this study will develop in vitro model
system to understanding mechanism of breast cancer
development as well as to help in developing specific
targeted therapies for breast cancer.
2.1. Collection and Processing of Human
Breast Cancer Tissue
Fresh specimen of human breast tumor was received
in sterile condition within an hour after surgery with
prier consent of patient as per the guidelines of ethical
committee of Jaslok Hospital and Research Center,
Mumbai, India and it was sent to the stem cell laboratory.
The tumor was cut into 2 mm pieces in sterile 1X Phos-
phate Buffer Saline (PBS) containing 1% Penstrep
(HiMedia, India) and washed 2 - 3 times. The cleaned
tissue fragments, then digested in 0.25% Trypsin-EDTA
(HiMedia, India) at 37˚C for 40 min. Tumor tissue ex-
plants were plated in 65 mm Nunc dishes which were
scratched with scarpel blade to adhere these tissue frag-
ments. The tissues pieces were fed with freshly prepared
DMEM (Dulbecco’s Modified Eagle’s Medium, HiMe-
dia, India) supplemented with 10% FBS (Fetal Bovine
Serum, Invitrogen, Carlsbad, CA), 1% Penicillin-strep-
tomycin, 1l/ml Insulin (Sigma, USA), 2 l/ml L-Gluta-
mine (HiMedia, India), 20 ng/ml EGF (Epidermal Growth
Factor, Sigma, USA) and dishes were incubated in a CO2
incubator at 37˚C with 5% CO2. Explants cultures were
observed daily under a phase contrast microscope for
outgrowth of cells from partially digested tissue frag-
ments. After 5th day many cells were seen outgrowing
from tissue, which were adhered to the bottom of the
petridish. These adhered cells were fed with fresh
growth media three times in a week. These adhered cells
started to multiply and reached confluent within 15 - 20
days. After attending confluency, the cells were passaged
using 0.25% Trypsin-EDTA and plated in new 50 mm
tissue culture flasks. The same procedure was repeated
after every confluency and cells were crypreserved at
2.2. Phase Contrast Microscop y
The morphology of cultured cells was observed using
phase contrast microscope equipped with TSview soft-
ware (Tucsen Imaging, Fuzhou, PR China) for observing
and capturing the images. Cells were regularly moni-
tored and images were captured for analysis.
2.3. Cryopreservation and Revival
Semi-confluent cultures of isolated breast cancer stem
cells were trypsinized with 0.25% Trypsin-EDTA for 3 -
5 minutes, centrifuged and washed with 1X PBS. The
cell pellet was suspended slowly in ready-made cell
freezing medium (Himedia, India) and transferred into
the cryovial and store at 85˚C. For revival, vials were
thawed and freezing medium was replaced by freshly
prepared DMEM medium. These cells maintained 90%
viability after revival.
2.4. Molecular Characterization of Breast
Cancer Stem Cells
Total RNA was extracted from tumor tissue of breast
cancer patient (In vivo study) and isolated breast cancer
stem cells (In vitro study) by using Trizol Reagent (Invi-
trogen, Carlsbad, CA). RNA was transcribed to cDNA
by using Applied Biosystems High Capacity cDNA Kit.
(Applied Biosystem, USA). The molecular markers
study was carried out by RT/PCR for Mesenchymal
phenotypes (CD105, CD13, CD73), Hematopoietic
phenotypes (CD34, CD45), Pluripotency and differenti-
ating markers (Oct4, Nanog, SOX2, LIF, Keratin18 and
Actin), Oncogenic markers (c-MYC, BCL2, COX2,
DAPK, EGFR and CD44), Make Singular - Cytokine
and Chemokine (IL6, TNF, SDF1 and CXCR4) and
BRCA2 gene mutation. PCR conditions and primer se-
P. Potdar et al. / Stem Cell Discovery 3 (2011) 21-28
Copyright © 2011 SciRes. http://www.scirp.org/journal/SCD/Openly accessible at
quences were described previously by Potdar and Sutar
2010 [24] whereas, sequences and their respective PCR
conditions for other than reported genes are listed in
Table 1. The PCR products were checked for their re-
spective amplification on 2% Agarose gel electrophore-
sis and photographed under UV light.
2.5. DNA Se quencing of BRCA2 Gene
Exon 11E of BRCA2 gene was amplified by PCR us-
ing the primer combination shown in Table 1. The PCR
product was sequenced using ABI 3100 Genetic analyzer
using BigDye Terminator reaction (Applied Biosystems,
Warrington, UK). The sequenced obtained were ana-
lyzed by using Bioedit software and the mutation ob-
tained was confirmed by repeating same sample for se-
quencing. Both sense and antisense strand sequencing
were carried out to confirm this mutation reported in this
3.1. Isolation of Human Breast Cancer
Mesenchymal Stem Cells (hBCMSCs)
from Human Non-Metastasis Breast
Cancer Tumor
The breast cancer stem cells were isolated and cul-
tured from non-metastasis breast cancer tumor is de-
scribed above. Culture dishes were regularly monitored
under phase contrast microscope for any out growth of
the breast cancer cells from breast cancer explants. Out-
growth of these cells was seen within 5 - 6 days of cul-
ture, from explant as shown in Figure 1(a). These cells
showed various morphological features from fibroblasts
like appearance to epithelial and endothelial cells like
appearance as shown in Figures 1(b)-(d). The cells
showed high rate of multiplication and observed many
mitotic cells. These cells became confluent within 20 -
25 days as shown in Figure 2(a). These cells were regu-
larly washed and fed with usual growth medium de-
scribed in material method. After confluency, cells were
passaged by trypsinisation with 0.25% PBS/Trypsin and
transferred into 50 mm culture flask. This process was
repeated several times after cell reached to confluency
and cryopreserved at different passages and kept at
–85˚C for future studies. Thus, this study has established
breast cancer stem cell line and was designated as human
Breast Cancer Mesenchymal Stem Cells (hBCMSCs) line
after confirming their phenotypes by molecular markers
described below. Presently, these cells are at passage 18
and still growing well (Figure 2(a)). It was also ob-
served that several cells developed mammosphere for-
mation and pluripotent clones as shown in Figures 2(b)
and (c).
Table 1. Shows sequences of primers used for respective molecular marker.
Primer Sequence 5’ 3’ Annealing () Size (bp)
TNF alpha Downstream GTTGACCTTGGTCTGGTAGG 58 555
SDF1 alpha Downstream CTTAGGGGATTTGGAAGTTT 57 188
P. Potdar et al. / Stem Cell Discovery 1 (2011) 21-28
Copyright © 2011 SciRes. Openl y accessible at http://www.scirp.org/journal/SCD/
(a) (b) (c) (d)
Figure 1. Shows human Breast Cancer Mesenchymal Stem Cells (hBCMSCs) derived from Breast cancer non- metastasis
tumor outgrowing from explants on 5th day of culture (a), Epithelial (b), Endothelial (c) and Fibroblast like hBCMSCs cells
(a) (b) (c)
Figure 2. Shows confluent culture of hBCMSCs cell line grown as monolayer cells (a), mammospheres formation (b) and
pluripotency clone (c).
3.2. Phase Contrast Microscop y of
hBCMSCs Cell Line
Phase contrast microscopy of hBCMSCs cell line was
shown in Figures 1(a)-(d) and Figures 2(a)-(c). There
are different cell types were seen emerging from ex-
plants confirming the heterogeneous population of breast
cancer cells associated with breast cancer development.
Figure 1(b) shown epithelial like cells emerging from
explants with epithelial sheet like structure having large
nuclei and dense cytoplasm whereas, Figure 1(c) shown
endothelial like cells, which are elongated and randomly
scatter cells with scanty cytoplasm having large nucleus
with 1 - 2 nucleoli. Figure 1(d) showed Geimsa stained
typical fibroblast like mesenchymal cells having scanty
cytoplasm, large nucleus and nucleoli with cytoplasmic
granules. Figure 2(a) showed a confluent culture with
elongated morphology. Figure 2(b) showed cells have
mammosphere formation during growth of these cells
whereas; Figure 2(c) showed formation of pluripotency
clone in growing culture.
3.3. Molecular Marker Analysis of Human
Breast Cancer Tumor and hBCMSCs
Cell Line
The present study was undertaken to find out molecu-
lar characterization of human breast cancer tumor (in
vivo) and hBCMSCs cell line (in vitro) to confirmed
specific phenotypes of these cells by using specific set of
stem cell markers such as mesenchymal stem cell mark-
ers, hematopoetic stem cell markers, pluripotency mark-
ers, oncogenic markers, cytokines and chemokines mar-
3.4. Selection of Positive Controls
RNA from CML, Normal blood cells and SCAT cell
line developed in our laboratory (24) were use as posi-
tive control for (CD105, CD45, CD34), (SOX2, Oct4,
Nanog, LIF, Keratin18, and DAPK) and (CD13, CD73,
COX2 and IL-6) respectively [24]. K562 cells were use
for positive control for c-MYC gene. Amplified PCR
products were sequenced for confirmation of required
P. Potdar et al. / Stem Cell Discovery 3 (2011) 21-28
Copyright © 2011 SciRes. Openl y accessible at http://www.scirp.org/journal/SCD/
gene sequences using specific primers. The sequences
obtained for each gene were confirmed from NCBI da-
tabase using BLAST software [data not shown] and
shown 100% match for these specific genes reported
here. On the basis of this, positive controls were selected
for this study.
3.4.1. Mesenchymal and Hema topoietic Markers
in Human Breast Tumor Cells and
hBCMSCs Cell Line
Mesenchymal and hematopoietic markers expression
in human breast tumor cells and hBCMSC cells is shown
in Figure 3. It was showed that tumor cells did not ex-
press CD105, CD13 and CD73 genes whereas, hBC-
MSCs cell line showed higher expression of all three
genes, thus confirming their mesenchymal stem cell
phenotypes. These cells also mildly expressed CD34 but
showed negative expression of CD45 gene indicating
that hBCMSCs cells may have hematopoetic phenotypes.
Both these genes were not expressed in tumor tissue.
3.4.2. Pluripotency Markers in Human Breast
Tumor Cells and hBCMSCs Cell Line
We have studied expression of Oct4, Nanog, SOX2,
LIF and Keratin18 genes as markers for pluripotency
and differentiation in tumor cells and in hBCMSCs cell
line respectively as shown in Figure 4. We have observed
that Oct 4, Nanog and SOX2 were expressed at in vivo
cells whereas, in hBCMSCs cell line Oct 4, Nanog were
mildly expressed and SOX2 was shown to be down reg-
ulated (Figure 4) indicate that SOX2 plays some role in
pathogenesis of Breast cancer and needs further investi-
gation. Differentiation markers LIF and keratin18 were
expressed equally in vivo and in vitro cells (Figure 4).
Figure 3. Shows expression of mes-
enchymal and hematopoietic stem
cell markers in Breast cancer tumor
tissue (in vivo) and in hBCMSCs cell
line (in vitro).
Figure 4. Shows expression of pluri-
potency stem cell markers in Breast
cancer tumor tissue (in vivo) and in
hBCMSCs cell line (in vitro).
3.4.3. Oncogenic Markers in Human Breast
Tumor Cells and hBCMSCs Cell Line
As hBCMSCs cell line was derived from non-metas-
tasis tumor cells, we thought of studying oncogenic
markers expression in these cells. We studied c-MYC,
COX2, BCL2, CD44, EGFR, and DAPK genes in tumor
as well as hBCMSCs cells as shown in Figure 5. It was
observed that c-MYC, EGFR, CD44, DAPK and BCL2
genes were expressed in hBCMSCs cell line whereas
only BCL2 and COX2 genes were expressed by tumor
tissue (Figure 5). Within these expressions, CD44 was
shown to be highly expressed in hBCMSCs cell line
confirming, the presence of initiating breast cancer tu-
mor cells. Expression of DAPK in tumor cells and
hBCMSCs cell line also confirmed their premalignant
status, as DAPK, a tumor suppressor gene expressed in
normal and initiated cells (Figure 5). These results were
further confirmed by studying chemokine receptor
CXCR4 expression in tumor and hBCMSCs cells. It was
also observed that CXCR4 gene was prominently ex-
pressed in tumor but mildly expressed in hBCMSCs cell
line indicating that the hBCMSCs cells isolated from
tumor were not metastatic but they are breast cancer
initiated cells. We also observed expression of SDF1 in
tumor and hBCMSCs cell line (Figure 6(b)). As cyto-
kines play very important role in mammary carcino-
genesis, we have studied expression of IL6 and TNF in
tumor and hBCMSCs cell line (Figure 6(a)). We ob-
served that TNF was expressed prominently in tumor
but shown mild expression in hBCMSCs cell line. Simi-
larly tumor tissue cells expressed mild expression of IL6
whereas, hBCMSCs cells showed significantly higher
expression of IL6 indicating specific role of IL6 in
breast carcinogenesis as reported by various investigator
P. Potdar et al. / Stem Cell Discovery 1 (2011) 21-28
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3.4.4. Expression of BRCA2 Gene Mutation in
Human Breast Tumor Cells and hBCMSCs
Cell Line
As our laboratory has established a founder mutation
in BRCA2 gene in Indian breast cancer patients [23], we
thought of evaluated this BRCA2 mutation in tumor
tissue as well as in hBCMSCs cell line by DNA se-
quencing (Figure 6(c)). We did find the mutation at exon
11E of BRCA 2 gene in tumor as well as hBCMSCs
cells indicating the cells isolated from tumor cells have
similar characteristic, as it appeared in vivo. Thus, we
can conclude that the cell line isolated from non-metas-
tasis breast cancer tumor is none other than Initiated
breast cancer stem cells causing tumor in humans.
Figure 5. Shows expression of oncogenic
cell markers in Breast cancer tumor tis-
sue (in vivo) and in hBCMSCs cell line
(in vitro).
Figure 6. (a) Shows expression of cytokine markers in
Breast cancer tumor tissue (in vivo) and in hBCMSCs cell
line (in vitro); (b) shows expression of chemokine markers
in Breast cancer tumor tissue (in vivo) and in hBCMSCs
cell line (in vitro); (c) shows electropherograms of exon 11E
of BRCA2 gene mutation (A3624G) observed in tumor
tissue as well as in hBCMSCs cell line.
Breast cancer is a complex disease, with multiple ge-
netic and environmental factors involved in its aetiology.
Recent advances in stem cell research have demon-
strated that the cancers originally develop from normal
cells, which gain the ability to proliferate aberrantly and
eventually turn malignant [1]. These malignant cells
have been defined as cancer stem cells. These cells have
exclusive ability to self-renewal and to differentiate into
the heterogeneous lineages of cancer cells that comprise
the tumor [1]. Two of major challenges in breast cancer
studies are 1) Isolation of Breast cancer stem cells from
breast tumor tissue and 2) identify breast cancer stem
cells using specific stem cell markers. The technology
for isolating Breast cancer stem cells from breast tumor
has been shown that CD24low/ CD44+ are a define popu-
lation of cells of potential breast cancer stem cells [5]. In
present study, we have examined non-metastasis breast
tumor for the presence of Breast cancer stem cells and
characterized them using molecular markers. We are the
first to isolate human breast cancer mesenchymal stem
cells from non-metastasis breast tumor. We have desig-
nated this cell line as human Breast Cancer Mesenchy-
mal Stem Cells (hBCMSCs), which is presently at pas-
sage 18. Recent studies have suggested the existence of
stem cells in the human mammary gland and shown
non-adherent mammospheres secretary cancer stem cells
with progenitor cell properties [12,13]. In our study, we
have observed similar phenomenon of formation of
mammospheres by hBCMSCs cells indicating clearly
that the cells isolated from non- metastatic tumor are
Breast cancer initiated stem cells.
Mesenchymal characteristic of these cells were con-
firmed by examination of CD105, CD13 and CD73
markers, which are putative markers for Mesenchymal
Stem Cells [14]. These cells also expressed hematopoi-
P. Potdar et al. / Stem Cell Discovery 3 (2011) 21-28
Copyright © 2011 SciRes. Openl y accessible at http://www.scirp.org/journal/SCD/
etic marker represented partial phenotype of hematopoi-
etic stem cells [15]. Shi et al. (2006) have shown that
transcription factors Oct-3/4, Nanog and SOX2 maintain
the expression of Pluripotent factors at a steady state [26]
whereas; Wilson et al. (1988) have shown that the LIF
maintains the developmental potential of embryonic
stem cells, which can be regarded as a pluripotency
marker [20]. In present study Breast cancer tumor cells
and hBCMSCs cells expressed Oct4, Nanog and LIF
showing their pluripotency. Expression of keratin 18 in
tumor cells and hBCMCSs cell line confirmed the
epithelial origin of these cells [27].
After establishment of breast cancer stem cell line, it
was very essential to know whether, the cell line pos-
sesses normal or malignant phenotypes so we have util-
ized oncogenic markers such as CD44, EGFR, c-MYC,
DAPK, COX2, BCL2, and SOX2 to study the malig-
nancy nature. Oncogenes overexpression is a common
phenomenon in the development and progression of
many human cancers hence it provide potential target for
cancer gene therapy [28]. Breast cancer cells were het-
erogeneous with respect to expression of a variety of cell
surface markers including CD44 and CD24. CD44 is a
complex transmembrane glycoprotein whose expression
is associated with drug resistance and metastatic pro-
gression in malignancy [21]. In present study, we have
shown that significant expression of CD44 in hBCMSCs
cell line and mild expression in tumor cells indicating
that the cells isolated from heterogeneous population of
tumor cells are breast cancer initiated cells. Studies have
shown that oncogenes c-MYC, COX2, EGFR and
DAPK plays important role in development of breast
cancer whereas, their overexpression or loss of expres-
sion is useful to study correlation with invasive potential
and metastasis in aggressive tumors and neoplastic tis-
sues[18,19,29,30]. Similarly expression of BCL2 and
SOX2 may be associated with metastasis, proliferation
and tumorigenesis of breast cancer cells [31,32]. Here in
the present study, there was significant expression of
c-MYC and EGFR in hBCMSCs cell line whereas, in
vivo study of tumor cells showed mild expression of
COX2 and down regulation of DAP Kinase gene sug-
gesting that the hBCMSCs cell lines contains initiated
breast cancer stem cells which may became transformed
to metastatic cancer cells when triggered by endogenous
Cytokines, such as IL-6 and Tumor Necrosis Factor
(TNFα), have an important role in regulating estrogen
synthesis and shown in vitro expression by aggressive
mesenchymal/basal-like breast cancer cell lines, tissues
and xenograft which promote malignant features in
breast cancer cells [25,33]. In present study, we found
low expression of IL6 in tumor cells whereas, signifi-
cantly high expression in hBCMSCs cell line which con-
firmed with the finding of other investigators [25,33].
Chemokine receptor CXCR4 showed significant high
expression in many solid tumors and also involved in
cell migration and invasion, as well as in angiogenesis
[34]. In present study, hBCMSCs cell line showed low
expression of CXCR4 indicating non-metastatic nature
of hBCMSCs cells isolated from tumor cells, which
showed significant high expression of CXCR4 gene.
We further thought of confirming cancer-initiated
cells by studying gene mutation in BRCA2 gene. Our
laboratory recently reported a specific founder mutation
in Exon 11E of BRCA2 gene in Indian population as an
early prognostic marker for Breast cancer [23]. In the
present study, we have sequenced DNA samples of
breast cancer tumor tissue and hBCMSCs cell line. Our
study showed that this founder mutation was at in vivo
and in vitro level indicating that the cells isolated from
non-metastasis breast cancer tumor are noting but Breast
cancer initiated cells which are responsible for develop-
ment of breast cancer.
Overall, this study establishes a technology for de-
velopment of breast cancer stem cell lines from
non-metastasis breast cancer tumor and represents an in
vitro model to understand the mechanism of Breast car-
cinogenesis. The identification of specific molecular
markers in these cells allows us to plan targeted thera-
pies for breast cancer.
We acknowledged with thanks the management of Jaslok Hospital
and Research Center, Mumbai, India for sanctioning and supporting
financial assistance for this stem cell research project No. 491. We are
also thankful to Dr. A. R. Khan, OT Manager for supplying us Breast
cancer Tumor tissues and Mrs. Naina Rane for technical help for this
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