Proliferation and Survival of Mammary Carcinoma Cells Are Influenced by Culture Conditions Used for Ex Vivo Expansion of CD34+Blood Progenitor Cells

Blood ◽  
1999 ◽  
Vol 93 (2) ◽  
pp. 746-755 ◽  
Author(s):  
A. Spyridonidis ◽  
W. Bernhardt ◽  
D. Behringer ◽  
G. Köhler ◽  
M. Azemar ◽  
...  
Keyword(s):  
Breast Cancer ◽  
Tumor Cells ◽  
Mammary Carcinoma ◽  
Ex Vivo ◽  
Culture Conditions ◽  
Ex Vivo Expansion ◽  
Carcinoma Cells ◽  
Serum Free ◽  
Cellular Contact ◽  
Mammary Carcinoma Cells

Abstract Malignant cell contamination in autologous transplants is a potential origin of tumor relapse. Ex vivo expansion of CD34+ blood progenitor cells (BPC) has been proposed as a tool to eliminate tumor cells from autografts. To characterize the influence of culture conditions on survival, growth, and clonogenicity of malignant cells, we isolated primary mammary carcinoma cells from pleural effusions and ascites of patients with metastatic breast cancer and cultured them in the presence of stem cell factor (SCF), interleukin-1β (IL-1β), IL-3, IL-6, and erythropoietin (EPO), ie, conditions previously shown to allow efficient ex vivo expansion of CD34+ BPC. In the presence of serum, tumor cells proliferated during a 7-day culture period and no significant growth-modulatory effect was attributable to the presence of hematopoietic growth factors. When transforming growth factor-β1 (TGF-β1) was added to these cultures, proliferation of breast cancer cells was reduced. Expansion of clonogenic tumor cells was seen in the presence of SCF + IL-1β + IL-3 + IL-6 + EPO, but was suppressed by TGF-β1. Cocultures of tumor cells in direct cellular contact with hematopoietic cells showed that tumor cell growth could be stimulated by ex vivo expanded hematopoietic cells at high cell densities (5 × 105/mL). In contrast, culture under serum-free conditions resulted in death of greater than 90% of breast cancer cells within 7 days and a further decrease in tumor cell numbers thereafter. In the serum-free cultures, hematopoietic cytokines and cellular contact with CD34+ BPC could not protect the tumor cells from death. Therefore, ex vivo expansion of CD34+ BPC in serum-free medium provides an environment for efficient purging of contaminating mammary carcinoma cells. These results have clinical significance for future protocols in autologous progenitor cell transplantation in cancer patients.

Proliferation and Survival of Mammary Carcinoma Cells Are Influenced by Culture Conditions Used for Ex Vivo Expansion of CD34+Blood Progenitor Cells

Blood ◽  
1999 ◽  
Vol 93 (2) ◽  
pp. 746-755 ◽  
Author(s):  
A. Spyridonidis ◽  
W. Bernhardt ◽  
D. Behringer ◽  
G. Köhler ◽  
M. Azemar ◽  
...  
Keyword(s):  
Breast Cancer ◽  
Tumor Cells ◽  
Mammary Carcinoma ◽  
Ex Vivo ◽  
Culture Conditions ◽  
Ex Vivo Expansion ◽  
Carcinoma Cells ◽  
Serum Free ◽  
Cellular Contact ◽  
Mammary Carcinoma Cells

Malignant cell contamination in autologous transplants is a potential origin of tumor relapse. Ex vivo expansion of CD34+ blood progenitor cells (BPC) has been proposed as a tool to eliminate tumor cells from autografts. To characterize the influence of culture conditions on survival, growth, and clonogenicity of malignant cells, we isolated primary mammary carcinoma cells from pleural effusions and ascites of patients with metastatic breast cancer and cultured them in the presence of stem cell factor (SCF), interleukin-1β (IL-1β), IL-3, IL-6, and erythropoietin (EPO), ie, conditions previously shown to allow efficient ex vivo expansion of CD34+ BPC. In the presence of serum, tumor cells proliferated during a 7-day culture period and no significant growth-modulatory effect was attributable to the presence of hematopoietic growth factors. When transforming growth factor-β1 (TGF-β1) was added to these cultures, proliferation of breast cancer cells was reduced. Expansion of clonogenic tumor cells was seen in the presence of SCF + IL-1β + IL-3 + IL-6 + EPO, but was suppressed by TGF-β1. Cocultures of tumor cells in direct cellular contact with hematopoietic cells showed that tumor cell growth could be stimulated by ex vivo expanded hematopoietic cells at high cell densities (5 × 105/mL). In contrast, culture under serum-free conditions resulted in death of greater than 90% of breast cancer cells within 7 days and a further decrease in tumor cell numbers thereafter. In the serum-free cultures, hematopoietic cytokines and cellular contact with CD34+ BPC could not protect the tumor cells from death. Therefore, ex vivo expansion of CD34+ BPC in serum-free medium provides an environment for efficient purging of contaminating mammary carcinoma cells. These results have clinical significance for future protocols in autologous progenitor cell transplantation in cancer patients.

scholarly journals Purging of Mammary Carcinoma Cells During Ex Vivo Culture of CD34+ Hematopoietic Progenitor Cells With Recombinant Immunotoxins

Blood ◽  
1998 ◽  
Vol 91 (5) ◽  
pp. 1820-1827 ◽  
Author(s):  
A. Spyridonidis ◽  
M. Schmidt ◽  
W. Bernhardt ◽  
A. Papadimitriou ◽  
M. Azemar ◽  
...  
Keyword(s):  
Progenitor Cells ◽  
Tumor Cells ◽  
Mammary Carcinoma ◽  
Ex Vivo ◽  
Ex Vivo Expansion ◽  
Carcinoma Cells ◽  
Hematopoietic Progenitor Cells ◽  
Egf Receptors ◽  
Hematopoietic Progenitor ◽  
Mammary Carcinoma Cells

Tumor cells have been found in autologous hematopoietic cell transplants used after high-dose chemotherapy. To specifically eliminate contaminating mammary tumor cells during ex vivo expansion of CD34+ hematopoietic progenitor cells, we used recombinant immunotoxins (ITs) directed against cell-surface antigens expressed on mammary carcinoma cells. ITs were expressed from fusion cDNAs combining a single-chain antibody fragment (scFv) directed against the Erb-B2 or epidermal growth factor (EGF) receptors with a truncatedPseudomonas exotoxin A fragment devoid of its cell-binding domain. CD34+ hematopoietic progenitor cells did not express Erb-B2 and EGF receptors as detected by Western blotting. Ex vivo expansion of total hematopoietic cells or of colony-forming cells from CD34+ progenitors in the presence of stem-cell factor (SCF), interleukin-1 (IL-1), IL-3, IL-6, and erythropoietin (Epo) was not affected when ITs were added to the cultures. In contrast, MDA-MB 453 and MCF-7 mammary carcinoma cells were depleted in a dose- and time-dependent manner by more than 3 log in coculture with CD34+ cells over a period of 7 days in the presence of 100 to 1,000 ng/mL of anti–Erb-B2 IT. This included elimination of the subpopulations with regrowth potential. Similarly, addition of either anti–Erb-B2 or anti-EGF receptor ITs to primary breast cancer cells isolated from patients with metastatic disease resulted in elimination of cytokeratin-positive cells in seven of seven samples. ITs are highly efficient and convenient to use for the depletion of mammary tumor cells during ex vivo expansion of hematopoietic progenitor-cell autografts.

scholarly journals Purging of Mammary Carcinoma Cells During Ex Vivo Culture of CD34+ Hematopoietic Progenitor Cells With Recombinant Immunotoxins

Blood ◽  
1998 ◽  
Vol 91 (5) ◽  
pp. 1820-1827 ◽  
Author(s):  
A. Spyridonidis ◽  
M. Schmidt ◽  
W. Bernhardt ◽  
A. Papadimitriou ◽  
M. Azemar ◽  
...  
Keyword(s):  
Progenitor Cells ◽  
Tumor Cells ◽  
Mammary Carcinoma ◽  
Ex Vivo ◽  
Ex Vivo Expansion ◽  
Carcinoma Cells ◽  
Hematopoietic Progenitor Cells ◽  
Egf Receptors ◽  
Hematopoietic Progenitor ◽  
Mammary Carcinoma Cells

Abstract Tumor cells have been found in autologous hematopoietic cell transplants used after high-dose chemotherapy. To specifically eliminate contaminating mammary tumor cells during ex vivo expansion of CD34+ hematopoietic progenitor cells, we used recombinant immunotoxins (ITs) directed against cell-surface antigens expressed on mammary carcinoma cells. ITs were expressed from fusion cDNAs combining a single-chain antibody fragment (scFv) directed against the Erb-B2 or epidermal growth factor (EGF) receptors with a truncatedPseudomonas exotoxin A fragment devoid of its cell-binding domain. CD34+ hematopoietic progenitor cells did not express Erb-B2 and EGF receptors as detected by Western blotting. Ex vivo expansion of total hematopoietic cells or of colony-forming cells from CD34+ progenitors in the presence of stem-cell factor (SCF), interleukin-1 (IL-1), IL-3, IL-6, and erythropoietin (Epo) was not affected when ITs were added to the cultures. In contrast, MDA-MB 453 and MCF-7 mammary carcinoma cells were depleted in a dose- and time-dependent manner by more than 3 log in coculture with CD34+ cells over a period of 7 days in the presence of 100 to 1,000 ng/mL of anti–Erb-B2 IT. This included elimination of the subpopulations with regrowth potential. Similarly, addition of either anti–Erb-B2 or anti-EGF receptor ITs to primary breast cancer cells isolated from patients with metastatic disease resulted in elimination of cytokeratin-positive cells in seven of seven samples. ITs are highly efficient and convenient to use for the depletion of mammary tumor cells during ex vivo expansion of hematopoietic progenitor-cell autografts.

scholarly journals Detection of Isolated Mammary Carcinoma Cells in Marrow of Patients with Primary Breast Cancer

1983 ◽  
Vol 76 (5) ◽  
pp. 359-364 ◽  
Author(s):  
D P Dearnaley ◽  
J P Sloane ◽  
S Imrie ◽  
R C Coombes ◽  
M G Ormerod ◽  
...  
Keyword(s):  
Breast Cancer ◽  
Bone Marrow ◽  
Bone Metastases ◽  
Mammary Carcinoma ◽  
Primary Breast Cancer ◽  
Epithelial Membrane Antigen ◽  
Single Cells ◽  
Carcinoma Cells ◽  
Mammary Carcinoma Cells

Single cells from mammary carcinoma infiltrating bone marrow can be detected in marrow aspirates using immunocytochemical stains for epithelial membrane antigen (EMA). This technique has been used to examine marrow aspirates taken from multiple sites from 24 patients at surgery for breast cancer. Ten of these patients had EMA-positive cells in their marrow, while 32 marrow samples from patients who did not have carcinoma were negative. These results have been combined with those obtained by taking aspirates from single sites from 47 breast patients without known skeletal deposits. Follow up showed that the patients with EMA-positive cells in their marrow developed bone metastases at a significantly faster rate.

BREAST CARCINOMAS SYNTHESIZE FACTORS WHICH INFLUENCE OSTEOBLAST-LIKE CELLS INDEPENDENTLY OF OSTEOCLASTS IN VITRO

1991 ◽  
Vol 128 (3) ◽  
pp. R5-R8 ◽  
Author(s):  
C E Evans ◽  
C Ward ◽  
I P Braidman
Keyword(s):  
Breast Cancer ◽  
Dna Synthesis ◽  
Mammary Carcinoma ◽  
Carcinoma Cells ◽  
Collagen Gels ◽  
Breast Carcinomas ◽  
Human Breast ◽  
Human Breast Carcinomas ◽  
Mammary Carcinoma Cells

ABSTRACT Bone metastases in breast cancer may be osteolytic, osteosclerotic, or a mixture of the two. Although stimulation of bone resorption by breast cancer cells has attracted some interest, the formation of osteosclerotic secondary tumours and the influence of human mammary carcinoma cells on osteoblasts (bone forming cells), both important in understanding breast cancer - bone interactions, have been largely neglected. We therefore examined the effects of conditioned medium (CM) from two cultured human breast cancer cell lines (MCF7 and ZR-75) and from primary cultures of breast carcinomas from two patients, on osteoblasts and recruitment of bone-resorbing cells (osteoclasts) in vitro. Osteoblast-like cells (BDC) were cultured from human trabecular bone explants. Osteoclast maturation was studied in fetal rat calvaria cultured on collagen gels. CM from the MCF-7 line and cells derived from one patient each inhibited BDC DNA synthesis, but stimulated osteoclast recruitment. In contrast, CM from the second patient's cells or ZR-75 enhanced DNA synthesis in BDC, but blocked osteoclast maturation. This suggests that human breast carcinomas secrete soluble factors which influence both osteoclasts and osteoblasts. A further unexpected implication is that mammary carcinoma cells may cause local osteosclerosis by directly stimulating osteoblasts, rather than through raised bone turnover in metastases.

Histone Deacetylase Inhibitors Promote the Ex Vivo Expansion of Cord Blood CD34+ Cells in Serum Free Cultures Accompanied by the Upregulation of Pluripotency Genes

Blood ◽  
2012 ◽  
Vol 120 (21) ◽  
pp. 345-345 ◽  
Author(s):  
Pratima Chaurasia ◽  
David Gajzer ◽  
Michael Feldman ◽  
Ronald Hoffman
Keyword(s):  
Cord Blood ◽  
Ex Vivo ◽  
Culture Conditions ◽  
Ex Vivo Expansion ◽  
Class I ◽  
Aldh Activity ◽  
Serum Free ◽  
Functional Potential ◽  
Cd34 Cells ◽  
Pluripotency Genes

Abstract Abstract 345 Previously, we have reported that various chromatin modifying agents(CMA) can epigenetically alter HSC fate decisions resulting in increased numbers of cord blood(CB)- SCID repopulating cells(SRC) and/or erythroid progenitor cells depending upon the CMA utilized (Blood. 2011,117(17):4632–41). We have hypothesized that treatment of CB CD34+ cells with histone deacetyalse inhibitors(HDACIs) might expand the numbers of CB-SRC and that such cell products could be used as allogeneic stem cell grafts. We developed serum free(SF) and containing(SC) culture conditions and assessed the ability of several HDACIs including valproic acid(VPA), scriptaid(SCR) and CAY10433(C433) to expand CB-SRC Each of these HDACIs were shown to inhibit both class I and II HDACs. The addition of VPA to SF cultures led to a >6,000 fold expansion of CD34+ cells and >25,000-fold expansion of CD34+CD90+ cells as compared to primary cells(PC). By contrast, SC cultures resulted in only a 54-fold expansion of CD34+ cells and a 75-fold expansion of CD34+90+ cells as compared to PC in the presence of VPA.In order to examine the functional characteristics of HDACIs treated CB-CD34+ cells in SF cultures we evaluated aldehyde dehydrogenase activity(ALDH) in these expanded CD34+ cells. A far greater proportion of CD34+ cells were ALDH+ following treatment with VPA in SF cultures as compared to SC cultures (82.2±8.6%, versus 37.0±0.9%). By contrast incubation in the SC cultures induced the generation of greater proportion of ALDH-CD34+ cells as compared to SF cultures (27.0±5.0% versus 3.7±2.0%). These findings indicate that the inclusion of serum to such cultures favors HSC differentiation and eventual HSC depletion while SF conditions favor expansion of more primitive HPC. Purified ALDH+CD34+ cells from SF cultures supplemented with HDACIs contained far greater absolute numbers of BFU-E and CFU-Mix (8.4×107±6.7×107/CB collection) as compared to SC cultures (1.4×107±8.8×106/CB collection; One way ANOVA p=.02). These data indicate that based upon their degree of ALDH activity and functional potential different phenotypically and functionally defined subsets of CD34+ cells are generated in SF as compared to SC cultures. In order to further examine the effects of VPA on the pluripotency of expanded CD34+ cells we examined the expression of Sox2, Oct4 and Nanog in CD34+ cells re-isolated following 7 days of treatment with HDACIs in SF and SC cultures. RNA and protein expression levels of Sox2, Oct4 and Nanog genes were greater in cells generated in SF cultures supplemented with HDACIs than that observed in CD34+ cells cultured in SC. Sox2 and Oct4 bind to the promoter of Nanog and up-regulate Nanog expression. In the SF but not SC cultures VPA treatment resulted in the up-regulation of Nanog indicating the functionality of the Sox2/Oct4 interactions. We also documented using confocal microscopy the elevated expression of the proteins derived from these pluripotency genes both in the cytoplasm and nucleus of CD34+ cells re-isolated after a week of treatment with each of the HDACIs. The functional potential of the re-isolated CD34+ cells following a week of treatment with VPA in SF medium was evaluated by transplanting the expanded cells into NOD/SCID/gamma(c)null mice. SRC capable of multi-lineage hematopoietic differentiation were detected in the marrow of recipients of VPA treated grafts 13-weeks after transplantation (%CD45, 33.5±13.4% with VPA treated grafts versus 13.0±7.7% with grafts generated in the presence of CA). Bone marrow cells from primary recipients receiving HDACI treated CD34+ cells grafts were also capable of generating greater numbers of cells belonging to myeloid and lymphoid lineages(%CD45, 3.3±1.0%) as compared to grafts receiving CA(%CD45, 0.65±0.16%) 15 weeks after their transplantation into secondary recipients. These data indicate that inhibitors of class I and II HDACs under SF culture conditions provide an effective pharmacological approach which can be utilized to promote the ex vivo expansion of functional CB-SRC which are characterized by high ALDH activity and expression of a variety of pluripotency genes. These findings indicate the importance of epigenetic events in determining phenotype and function of dividing SRC in vitro. Disclosures: Hoffman: NYSTEM: Research Funding.

Effects of Salvia officinalis L. on 7, 12-dimethylbenz[a]anthracene-induced breast cancer in rats and mouse mammary carcinoma cells (4T1)

2017 ◽  
Vol 26 (5) ◽  
pp. 1003-1015
Author(s):  
Malihezaman Monsefi ◽  
Zahra Azarbahram ◽  
Mehrnaz Abedian ◽  
Sara Behrozimoghadam ◽  
Mohammad Javad Ashraf
Keyword(s):  
Breast Cancer ◽  
Mammary Carcinoma ◽  
Carcinoma Cells ◽  
Salvia Officinalis ◽  
Mouse Mammary Carcinoma ◽  
Mammary Carcinoma Cells ◽  
Salvia Officinalis L

scholarly journals Canine Metastatic Mammary Carcinoma Cells in Smears from Genital Epithelium

1974 ◽  
Vol 11 (1) ◽  
pp. 20-28 ◽  
Author(s):  
J. F. Roszel
Keyword(s):  
Tumor Cells ◽  
Mammary Carcinoma ◽  
Primary Lesion ◽  
Postmortem Examination ◽  
Carcinoma Cells ◽  
Tissue Sections ◽  
Mammary Carcinoma Cells

Smears from the genital epithelium of five dogs with mammary carcinoma contained cells that morphologically resembled those in tissue sections of the primary lesion. During postmortem examination of three of these animals cells exfoliated from selected sites were examined to identify the probable area of desquamation of tumor cells. Histologic sections of the grossly normal vestibules contained submucosal micrometastatic foci of tumor.

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