Lisa Arendt - Academia.edu (original) (raw)

Papers by Lisa Arendt

Research paper thumbnail of Prolactin induces ERα-positive and ERα-negative mammary cancer in transgenic mice

Oncogene, Jul 24, 2003

The role of prolactin in human breast cancer has been controversial. However, it is now apparent ... more The role of prolactin in human breast cancer has been controversial. However, it is now apparent that human mammary epithelial cells can synthesize prolactin endogenously, permitting autocrine/paracrine actions within the mammary gland that are independent of pituitary prolactin. To model this local mammary production of prolactin (PRL), we have generated mice that overexpress prolactin within mammary epithelial cells under the control of a hormonally nonresponsive promoter, neurelated lipocalin (NRL). In each of the two examined NRL-PRL transgenic mouse lineages, female virgin mice display mammary developmental abnormalities, mammary intraepithelial neoplasias, and invasive neoplasms. Prolactin increases proliferation in morphologically normal alveoli and ducts, as well as in lesions. The tumors are of varied histotype, but papillary adenocarcinomas and adenosquamous neoplasms predominate. Neoplasms can be separated into two populations: one is estrogen receptor alpha (ERa) positive (greater than 15% of the cells stain for ERa), and the other is ERaÀ (o3%). ERa expression does not correlate with tumor histotype, or proliferative or apoptotic indices. These studies provide a mouse model of hormonally dependent breast cancer, and, perhaps most strikingly, a model in which some neoplasms retain ERa, as occurs in the human disease.

Research paper thumbnail of Peripheral Serotonin Ablation Affects Cyclins and Cellular Proliferation during Parturition, but not late Pregnancy in Dams Fed a High-Fat Diet

Primer sequences used in a short communication which investigates the effect of serotonin ablatio... more Primer sequences used in a short communication which investigates the effect of serotonin ablation on cell cycle genes and cellular proliferation during late gestation and parturition in the mammary gland.

Research paper thumbnail of Additional file 5: Figure S5. of Obesity reversibly depletes the basal cell population and enhances mammary epithelial cell estrogen receptor alpha expression and progenitor activity

Obesity reduces myoepithelial cells and enhances ERα-positive luminal cells in mice fed a high-fa... more Obesity reduces myoepithelial cells and enhances ERα-positive luminal cells in mice fed a high-fat diet (HFD) during puberty. A The 3-week-old C57Bl/6 female mice were started on a control diet (Con) or HFD for 17 weeks. Mammary epithelial cells were isolated and quantified using flow cytometry as described in "Methods". The ratio of luminal cells (EpCAMhiCD49flo) to basal cells (EpCAMloCD49fhi) was quantified in three experiments (n = 6 mice). The percentage of luminal and basal cells was determined from total mammary lineage positive cells. B Myoepithelial cells were detected using immunofluorescence for SMA within the mammary glands of C57Bl/6 and FVB/N mice fed HFD or Con starting at 3 weeks of age. SMA continuity was scored as described in "Methods" (n = 3 images/gland; 5 mice/group). C ERα+ luminal cells were immunohistochemically detected within the mammary glands of C57Bl/6 and FVB/N mice fed the HFD or Con starting at 3 weeks of age (n = 5 images/gland; ...

Research paper thumbnail of Additional file 4: Figure S4. of Obesity reversibly depletes the basal cell population and enhances mammary epithelial cell estrogen receptor alpha expression and progenitor activity

Effect of obesity on estrus cycle in C57Bl/6 and FVB/N mice. A Quantification of average days in ... more Effect of obesity on estrus cycle in C57Bl/6 and FVB/N mice. A Quantification of average days in estrus and diestrus over 2 weeks using vaginal cytology from C57Bl/6 fed a high-fat diet (HFD) or control diet (Con) starting at 3 weeks of age (n = 9/group). B Quantification of average days in estrus and diestrus over 2 weeks using vaginal cytology from FVB/N mice fed a HFD or Con starting at 3 weeks of age (n = 9/group). C Uterine weights after 17 weeks on HFD or Con from C57Bl/6 mice fed at 3 weeks of age. D Uterine weights after 17 weeks on HFD or Con from FVB/N fed at 3 weeks of age. Bars represent mean ± s.d. (PDF 97 kb)

Research paper thumbnail of Additional file 3: Figure S3. of Obesity reversibly depletes the basal cell population and enhances mammary epithelial cell estrogen receptor alpha expression and progenitor activity

Obesogenesis in response to a high-fat diet with respect to timing and mouse strain. A Eight-week... more Obesogenesis in response to a high-fat diet with respect to timing and mouse strain. A Eight-week-old C57Bl/6 mice (B6-8 wk) were fed a high-fat diet (HFD) or control diet (Con). Weight gain was determined for 17 weeks (n = 7 mice/group; mean ± s.e.m.). Differences were determined using two way ANOVA. Eight-week-old FVB/N female mice (FVB-8 wk) were fed a HFD or Con for 14 weeks (n = 5 mice/group). Body weight (B) and weight gain (C) were measured (mean ± s.e.m.). No differences were detected using two way ANOVA. D Inguinal gland weight from FVB-8 wk mice fed either HFD or Con. E Quantification of tertiary branching in whole mounts from HFD and Con inguinal mammary glands from FVB-8 wk mice. F Three-week-old C57Bl/6 mice (B6-3 wk) were fed a HFD or Con. Weight gain was determined for 17 weeks (n = 9 mice/group; mean ± s.e.m.). G Fat surrounding the uterus (gonadal fat) was weighed for B6-3 wk Con and Mice fed HFD (n = 9 mice/group). H Quantification of adipocyte diameters from mamma...

Research paper thumbnail of Additional file 2: Figure S2. of Obesity reversibly depletes the basal cell population and enhances mammary epithelial cell estrogen receptor alpha expression and progenitor activity

Flow cytometry and cytokeratin 14 immunofluorescence staining. A Representative contour plots to ... more Flow cytometry and cytokeratin 14 immunofluorescence staining. A Representative contour plots to demonstrate gating for mouse mammary epithelial cell flow cytometry. Cells were gated in SSC and FSC to eliminate debris. Live cells were identified as negative for fixable viability dye eFluor 780. PE-conjugated stromal lineage markers, including CD31, CD45, and Ter119, were then removed from further analysis through gating. B Representative contour plots to demonstrate gating for human breast epithelial cell flow cytometry. Human breast epithelial cells were dissociated and lineage-depleted using bead-conjugated antibodies prior to analysis for flow cytometry as described in "Methods". Cells were gated in SSC and FSC to eliminate debris, and live cells were identified as negative for fixable viability dye eFluor 780. C Cytokeratin (CK)14-positive cells (red) were detected in human reduction mammoplasty tissue using immunofluorescence. Nuclei were detected with DAPI (blue). CK...

Research paper thumbnail of Additional file 1: Figure S1. of Obesity reversibly depletes the basal cell population and enhances mammary epithelial cell estrogen receptor alpha expression and progenitor activity

High-fat diet (HFD) feeding increases mammary adipocyte size and inflammation. A C57Bl/6 female m... more High-fat diet (HFD) feeding increases mammary adipocyte size and inflammation. A C57Bl/6 female mice were fed a HFD or control diet (Con) starting at 8 weeks of age. Fat surrounding the uterus (gonadal fat) was weighed in Con and HFD mice (n = 7 mice/group). B Representative images of mammary glands from Con and HFD mice, stained with F4/80 to detect crown-like structures. C Quantification of adipocyte diameters from mammary glands of Con and HFD mice (n = 7 mice/group). D Quantification of crown-like structures from mammary glands of Con and HFD mice (n = 7 mice/group). No crown-like structures were observed in the mammary glands of Con mice. Bars represent mean ± s.d. Magnification bar = 100 μm. (PDF 230 kb)

Research paper thumbnail of Divide and Conquer: Isolating Cell Populations to Investigate How Breast Cancer Risk Factors Alter the Breast Microenvironment

Methods in Molecular Biology, 2022

Breast cancer is a multifactorial disease with risk factors that are fixed or modifiable. Underst... more Breast cancer is a multifactorial disease with risk factors that are fixed or modifiable. Understanding how these risk factors interact within breast tissue may provide insight into how to improve interventions or chemoprevention strategies to reduce breast cancer incidence. Here we describe methods to utilize breast tissue from patients with defined risk factors undergoing reduction mammoplasty or prophylactic mastectomy to isolate epithelial cells, stromal cells, adipocytes, and macrophages to investigate how risk factors impact distinct cell populations within breast tissue. Following enzymatic digestion of breast tissue, adipocyte-enriched, stromal cell, and epithelial organoid fractions can be isolated. Using antibody-conjugated beads, further cell populations, such as macrophages, can be isolated for molecular analysis. These methods can be adapted to sequentially isolate other cell populations based on specific cell surface markers and are useful for small-sized breast tissue specimens.

Research paper thumbnail of Additional file 1: of Growth of human breast tissues from patient cells in 3D hydrogel scaffolds

Supporting Methods [ 22 , 23 , 50 ]. (DOC 66 kb)

Research paper thumbnail of Additional file 6: Movie S4. of Growth of human breast tissues from patient cells in 3D hydrogel scaffolds

Time-lapse microscopy depicting dynamic cellular migration in a growing organoid. The arrows indi... more Time-lapse microscopy depicting dynamic cellular migration in a growing organoid. The arrows indicate the direction of mass migration away from the leading edge of one duct and into another elongating duct. At the outset of the movie, the organoid had been grown for 8Â days. Images were captured every 10Â minutes. Frame rate: eight frames per second. (AVI 7540 kb)

Research paper thumbnail of Additional file 4: Movie S2. of Growth of human breast tissues from patient cells in 3D hydrogel scaffolds

Time-lapse microscopy depicting a rotating lobule and the behavior of leader cells. The arrow ind... more Time-lapse microscopy depicting a rotating lobule and the behavior of leader cells. The arrow indicates the direction of rotation of the lobule on the left. Arrowheads indicate the location of leader cells extending from two elongating outgrowths. At the outset of the movie, the organoid had been grown for 8Â days. Images were captured every 10Â minutes. Frame rate: eight frames per second. (AVI 9677 kb)

Research paper thumbnail of Additional file 7: Movie S5. of Growth of human breast tissues from patient cells in 3D hydrogel scaffolds

Twenty-four-hour time-lapse movie of an organoid growing in hydrogel. Still frames from this movi... more Twenty-four-hour time-lapse movie of an organoid growing in hydrogel. Still frames from this movie are depicted in Fig. 4d. At the outset of the movie, the organoid had been grown for 8 days. Images were captured every 10 minutes. Frame rate: eight frames per second. (AVI 19903 kb)

Research paper thumbnail of Additional file 3: Movie S1. of Growth of human breast tissues from patient cells in 3D hydrogel scaffolds

Confocal Z-series panning through the duct depicted in Fig. 2c (right). Z fields are separated by... more Confocal Z-series panning through the duct depicted in Fig. 2c (right). Z fields are separated by 1.11 μm, spanning a total of 71.94 μm. Note that the exterior of the structure is completely encapsulated by the myoepithelial layer of cells (CK14, green) and that the luminal layer is restricted to the inner layer of cells (CK8/18, red). (AVI 202753 kb)

Research paper thumbnail of Additional file 8: Movie S6. of Growth of human breast tissues from patient cells in 3D hydrogel scaffolds

Time-lapse microscopy depicting a leader cell redirecting the orientation of elongation of an out... more Time-lapse microscopy depicting a leader cell redirecting the orientation of elongation of an outgrowth. At the outset of the movie, the organoid had been grown for 8Â days. Images were captured every 10Â minutes. Frame rate: eight frames per second. (AVI 15328 kb)

Research paper thumbnail of Additional file 5: Movie S3. of Growth of human breast tissues from patient cells in 3D hydrogel scaffolds

Time-lapse microscopy depicting dynamic cellular migration in a growing organoid. The arrows indi... more Time-lapse microscopy depicting dynamic cellular migration in a growing organoid. The arrows indicate the direction of mass migration along two elongating ducts from the core of the organoid toward the leading edge of the ducts. At the outset of the movie, the organoid had been grown for 8Â days. Images were captured every 10Â minutes. Frame rate: eight frames per second. (AVI 5319 kb)

Research paper thumbnail of Additional file 2: Supplemental Figures. of Growth of human breast tissues from patient cells in 3D hydrogel scaffolds

Figure S1 Schematic for the dissociation of reduction mammoplasty tissue. Schematic representatio... more Figure S1 Schematic for the dissociation of reduction mammoplasty tissue. Schematic representation of tissue dissociation and purification of epithelium. For more details, see Additional file 1: Supporting Methods. Figure S2 Physical characterization of collagen and ECM hydrogels. a Young's modulus was measured at least three times for each of three independent replicates (red, blue, and green) for collagen gels and ECM hydrogels. Plotted are the mean and standard deviation for the replicates. b The swelling ratio was calculated for collagen gels and ECM hydrogels for four independent replicates. Plotted are the mean and standard deviation. *p

Research paper thumbnail of WNT enhances acinar progenitor activity through LRP6 signaling

Research paper thumbnail of Growth of human breast tissues from patient cells in 3D hydrogel scaffolds

Breast cancer research : BCR, 2016

Three-dimensional (3D) cultures have proven invaluable for expanding human tissues for basic rese... more Three-dimensional (3D) cultures have proven invaluable for expanding human tissues for basic research and clinical applications. In both contexts, 3D cultures are most useful when they (1) support the outgrowth of tissues from primary human cells that have not been immortalized through extensive culture or viral infection and (2) include defined, physiologically relevant components. Here we describe a 3D culture system with both of these properties that stimulates the outgrowth of morphologically complex and hormone-responsive mammary tissues from primary human breast epithelial cells. Primary human breast epithelial cells isolated from patient reduction mammoplasty tissues were seeded into 3D hydrogels. The hydrogel scaffolds were composed of extracellular proteins and carbohydrates present in human breast tissue and were cultured in serum-free medium containing only defined components. The physical properties of these hydrogels were determined using atomic force microscopy. Tissue...

Research paper thumbnail of Breast cancer microenvironment and obesity: challenges for therapy

Cancer and Metastasis Reviews

Women with obesity who develop breast cancer have a worsened prognosis with diminished survival r... more Women with obesity who develop breast cancer have a worsened prognosis with diminished survival rates and increased rates of metastasis. Obesity is also associated with decreased breast cancer response to endocrine and chemotherapeutic treatments. Studies utilizing multiple in vivo models of obesity as well as human breast tumors have enhanced our understanding of how obesity alters the breast tumor microenvironment. Changes in the complement and function of adipocytes, adipose-derived stromal cells, immune cells, and endothelial cells and remodeling of the extracellular matrix all contribute to the rapid growth of breast tumors in the context of obesity. Interactions of these cells enhance secretion of cytokines and adipokines as well as local levels of estrogen within the breast tumor microenvironment that promote resistance to multiple therapies. In this review, we will discuss our current understanding of the impact of obesity on the breast tumor microenvironment, how obesity-in...

Research paper thumbnail of Obesity-Activated Lung Stromal Cells Promote Myeloid Lineage Cell Accumulation and Breast Cancer Metastasis

Cancers

Obesity is correlated with increased incidence of breast cancer metastasis; however, the mechanis... more Obesity is correlated with increased incidence of breast cancer metastasis; however, the mechanisms underlying how obesity promotes metastasis are unclear. In a diet-induced obese mouse model, obesity enhanced lung metastasis in both the presence and absence of primary mammary tumors and increased recruitment of myeloid lineage cells into the lungs. In the absence of tumors, obese mice demonstrated increased numbers of myeloid lineage cells and elevated collagen fibers within the lung stroma, reminiscent of premetastatic niches formed by primary tumors. Lung stromal cells isolated from obese tumor-naïve mice showed increased proliferation, contractility, and expression of extracellular matrix, inflammatory markers and transforming growth factor beta-1 (TGFβ1). Conditioned media from lung stromal cells from obese mice promoted myeloid lineage cell migration in vitro in response to colony-stimulating factor 2 (CSF2) expression and enhanced invasion of tumor cells. Together, these resu...

Research paper thumbnail of Prolactin induces ERα-positive and ERα-negative mammary cancer in transgenic mice

Oncogene, Jul 24, 2003

The role of prolactin in human breast cancer has been controversial. However, it is now apparent ... more The role of prolactin in human breast cancer has been controversial. However, it is now apparent that human mammary epithelial cells can synthesize prolactin endogenously, permitting autocrine/paracrine actions within the mammary gland that are independent of pituitary prolactin. To model this local mammary production of prolactin (PRL), we have generated mice that overexpress prolactin within mammary epithelial cells under the control of a hormonally nonresponsive promoter, neurelated lipocalin (NRL). In each of the two examined NRL-PRL transgenic mouse lineages, female virgin mice display mammary developmental abnormalities, mammary intraepithelial neoplasias, and invasive neoplasms. Prolactin increases proliferation in morphologically normal alveoli and ducts, as well as in lesions. The tumors are of varied histotype, but papillary adenocarcinomas and adenosquamous neoplasms predominate. Neoplasms can be separated into two populations: one is estrogen receptor alpha (ERa) positive (greater than 15% of the cells stain for ERa), and the other is ERaÀ (o3%). ERa expression does not correlate with tumor histotype, or proliferative or apoptotic indices. These studies provide a mouse model of hormonally dependent breast cancer, and, perhaps most strikingly, a model in which some neoplasms retain ERa, as occurs in the human disease.

Research paper thumbnail of Peripheral Serotonin Ablation Affects Cyclins and Cellular Proliferation during Parturition, but not late Pregnancy in Dams Fed a High-Fat Diet

Primer sequences used in a short communication which investigates the effect of serotonin ablatio... more Primer sequences used in a short communication which investigates the effect of serotonin ablation on cell cycle genes and cellular proliferation during late gestation and parturition in the mammary gland.

Research paper thumbnail of Additional file 5: Figure S5. of Obesity reversibly depletes the basal cell population and enhances mammary epithelial cell estrogen receptor alpha expression and progenitor activity

Obesity reduces myoepithelial cells and enhances ERα-positive luminal cells in mice fed a high-fa... more Obesity reduces myoepithelial cells and enhances ERα-positive luminal cells in mice fed a high-fat diet (HFD) during puberty. A The 3-week-old C57Bl/6 female mice were started on a control diet (Con) or HFD for 17 weeks. Mammary epithelial cells were isolated and quantified using flow cytometry as described in "Methods". The ratio of luminal cells (EpCAMhiCD49flo) to basal cells (EpCAMloCD49fhi) was quantified in three experiments (n = 6 mice). The percentage of luminal and basal cells was determined from total mammary lineage positive cells. B Myoepithelial cells were detected using immunofluorescence for SMA within the mammary glands of C57Bl/6 and FVB/N mice fed HFD or Con starting at 3 weeks of age. SMA continuity was scored as described in "Methods" (n = 3 images/gland; 5 mice/group). C ERα+ luminal cells were immunohistochemically detected within the mammary glands of C57Bl/6 and FVB/N mice fed the HFD or Con starting at 3 weeks of age (n = 5 images/gland; ...

Research paper thumbnail of Additional file 4: Figure S4. of Obesity reversibly depletes the basal cell population and enhances mammary epithelial cell estrogen receptor alpha expression and progenitor activity

Effect of obesity on estrus cycle in C57Bl/6 and FVB/N mice. A Quantification of average days in ... more Effect of obesity on estrus cycle in C57Bl/6 and FVB/N mice. A Quantification of average days in estrus and diestrus over 2 weeks using vaginal cytology from C57Bl/6 fed a high-fat diet (HFD) or control diet (Con) starting at 3 weeks of age (n = 9/group). B Quantification of average days in estrus and diestrus over 2 weeks using vaginal cytology from FVB/N mice fed a HFD or Con starting at 3 weeks of age (n = 9/group). C Uterine weights after 17 weeks on HFD or Con from C57Bl/6 mice fed at 3 weeks of age. D Uterine weights after 17 weeks on HFD or Con from FVB/N fed at 3 weeks of age. Bars represent mean ± s.d. (PDF 97 kb)

Research paper thumbnail of Additional file 3: Figure S3. of Obesity reversibly depletes the basal cell population and enhances mammary epithelial cell estrogen receptor alpha expression and progenitor activity

Obesogenesis in response to a high-fat diet with respect to timing and mouse strain. A Eight-week... more Obesogenesis in response to a high-fat diet with respect to timing and mouse strain. A Eight-week-old C57Bl/6 mice (B6-8 wk) were fed a high-fat diet (HFD) or control diet (Con). Weight gain was determined for 17 weeks (n = 7 mice/group; mean ± s.e.m.). Differences were determined using two way ANOVA. Eight-week-old FVB/N female mice (FVB-8 wk) were fed a HFD or Con for 14 weeks (n = 5 mice/group). Body weight (B) and weight gain (C) were measured (mean ± s.e.m.). No differences were detected using two way ANOVA. D Inguinal gland weight from FVB-8 wk mice fed either HFD or Con. E Quantification of tertiary branching in whole mounts from HFD and Con inguinal mammary glands from FVB-8 wk mice. F Three-week-old C57Bl/6 mice (B6-3 wk) were fed a HFD or Con. Weight gain was determined for 17 weeks (n = 9 mice/group; mean ± s.e.m.). G Fat surrounding the uterus (gonadal fat) was weighed for B6-3 wk Con and Mice fed HFD (n = 9 mice/group). H Quantification of adipocyte diameters from mamma...

Research paper thumbnail of Additional file 2: Figure S2. of Obesity reversibly depletes the basal cell population and enhances mammary epithelial cell estrogen receptor alpha expression and progenitor activity

Flow cytometry and cytokeratin 14 immunofluorescence staining. A Representative contour plots to ... more Flow cytometry and cytokeratin 14 immunofluorescence staining. A Representative contour plots to demonstrate gating for mouse mammary epithelial cell flow cytometry. Cells were gated in SSC and FSC to eliminate debris. Live cells were identified as negative for fixable viability dye eFluor 780. PE-conjugated stromal lineage markers, including CD31, CD45, and Ter119, were then removed from further analysis through gating. B Representative contour plots to demonstrate gating for human breast epithelial cell flow cytometry. Human breast epithelial cells were dissociated and lineage-depleted using bead-conjugated antibodies prior to analysis for flow cytometry as described in "Methods". Cells were gated in SSC and FSC to eliminate debris, and live cells were identified as negative for fixable viability dye eFluor 780. C Cytokeratin (CK)14-positive cells (red) were detected in human reduction mammoplasty tissue using immunofluorescence. Nuclei were detected with DAPI (blue). CK...

Research paper thumbnail of Additional file 1: Figure S1. of Obesity reversibly depletes the basal cell population and enhances mammary epithelial cell estrogen receptor alpha expression and progenitor activity

High-fat diet (HFD) feeding increases mammary adipocyte size and inflammation. A C57Bl/6 female m... more High-fat diet (HFD) feeding increases mammary adipocyte size and inflammation. A C57Bl/6 female mice were fed a HFD or control diet (Con) starting at 8 weeks of age. Fat surrounding the uterus (gonadal fat) was weighed in Con and HFD mice (n = 7 mice/group). B Representative images of mammary glands from Con and HFD mice, stained with F4/80 to detect crown-like structures. C Quantification of adipocyte diameters from mammary glands of Con and HFD mice (n = 7 mice/group). D Quantification of crown-like structures from mammary glands of Con and HFD mice (n = 7 mice/group). No crown-like structures were observed in the mammary glands of Con mice. Bars represent mean ± s.d. Magnification bar = 100 μm. (PDF 230 kb)

Research paper thumbnail of Divide and Conquer: Isolating Cell Populations to Investigate How Breast Cancer Risk Factors Alter the Breast Microenvironment

Methods in Molecular Biology, 2022

Breast cancer is a multifactorial disease with risk factors that are fixed or modifiable. Underst... more Breast cancer is a multifactorial disease with risk factors that are fixed or modifiable. Understanding how these risk factors interact within breast tissue may provide insight into how to improve interventions or chemoprevention strategies to reduce breast cancer incidence. Here we describe methods to utilize breast tissue from patients with defined risk factors undergoing reduction mammoplasty or prophylactic mastectomy to isolate epithelial cells, stromal cells, adipocytes, and macrophages to investigate how risk factors impact distinct cell populations within breast tissue. Following enzymatic digestion of breast tissue, adipocyte-enriched, stromal cell, and epithelial organoid fractions can be isolated. Using antibody-conjugated beads, further cell populations, such as macrophages, can be isolated for molecular analysis. These methods can be adapted to sequentially isolate other cell populations based on specific cell surface markers and are useful for small-sized breast tissue specimens.

Research paper thumbnail of Additional file 1: of Growth of human breast tissues from patient cells in 3D hydrogel scaffolds

Supporting Methods [ 22 , 23 , 50 ]. (DOC 66 kb)

Research paper thumbnail of Additional file 6: Movie S4. of Growth of human breast tissues from patient cells in 3D hydrogel scaffolds

Time-lapse microscopy depicting dynamic cellular migration in a growing organoid. The arrows indi... more Time-lapse microscopy depicting dynamic cellular migration in a growing organoid. The arrows indicate the direction of mass migration away from the leading edge of one duct and into another elongating duct. At the outset of the movie, the organoid had been grown for 8Â days. Images were captured every 10Â minutes. Frame rate: eight frames per second. (AVI 7540 kb)

Research paper thumbnail of Additional file 4: Movie S2. of Growth of human breast tissues from patient cells in 3D hydrogel scaffolds

Time-lapse microscopy depicting a rotating lobule and the behavior of leader cells. The arrow ind... more Time-lapse microscopy depicting a rotating lobule and the behavior of leader cells. The arrow indicates the direction of rotation of the lobule on the left. Arrowheads indicate the location of leader cells extending from two elongating outgrowths. At the outset of the movie, the organoid had been grown for 8Â days. Images were captured every 10Â minutes. Frame rate: eight frames per second. (AVI 9677 kb)

Research paper thumbnail of Additional file 7: Movie S5. of Growth of human breast tissues from patient cells in 3D hydrogel scaffolds

Twenty-four-hour time-lapse movie of an organoid growing in hydrogel. Still frames from this movi... more Twenty-four-hour time-lapse movie of an organoid growing in hydrogel. Still frames from this movie are depicted in Fig. 4d. At the outset of the movie, the organoid had been grown for 8 days. Images were captured every 10 minutes. Frame rate: eight frames per second. (AVI 19903 kb)

Research paper thumbnail of Additional file 3: Movie S1. of Growth of human breast tissues from patient cells in 3D hydrogel scaffolds

Confocal Z-series panning through the duct depicted in Fig. 2c (right). Z fields are separated by... more Confocal Z-series panning through the duct depicted in Fig. 2c (right). Z fields are separated by 1.11 μm, spanning a total of 71.94 μm. Note that the exterior of the structure is completely encapsulated by the myoepithelial layer of cells (CK14, green) and that the luminal layer is restricted to the inner layer of cells (CK8/18, red). (AVI 202753 kb)

Research paper thumbnail of Additional file 8: Movie S6. of Growth of human breast tissues from patient cells in 3D hydrogel scaffolds

Time-lapse microscopy depicting a leader cell redirecting the orientation of elongation of an out... more Time-lapse microscopy depicting a leader cell redirecting the orientation of elongation of an outgrowth. At the outset of the movie, the organoid had been grown for 8Â days. Images were captured every 10Â minutes. Frame rate: eight frames per second. (AVI 15328 kb)

Research paper thumbnail of Additional file 5: Movie S3. of Growth of human breast tissues from patient cells in 3D hydrogel scaffolds

Time-lapse microscopy depicting dynamic cellular migration in a growing organoid. The arrows indi... more Time-lapse microscopy depicting dynamic cellular migration in a growing organoid. The arrows indicate the direction of mass migration along two elongating ducts from the core of the organoid toward the leading edge of the ducts. At the outset of the movie, the organoid had been grown for 8Â days. Images were captured every 10Â minutes. Frame rate: eight frames per second. (AVI 5319 kb)

Research paper thumbnail of Additional file 2: Supplemental Figures. of Growth of human breast tissues from patient cells in 3D hydrogel scaffolds

Figure S1 Schematic for the dissociation of reduction mammoplasty tissue. Schematic representatio... more Figure S1 Schematic for the dissociation of reduction mammoplasty tissue. Schematic representation of tissue dissociation and purification of epithelium. For more details, see Additional file 1: Supporting Methods. Figure S2 Physical characterization of collagen and ECM hydrogels. a Young's modulus was measured at least three times for each of three independent replicates (red, blue, and green) for collagen gels and ECM hydrogels. Plotted are the mean and standard deviation for the replicates. b The swelling ratio was calculated for collagen gels and ECM hydrogels for four independent replicates. Plotted are the mean and standard deviation. *p

Research paper thumbnail of WNT enhances acinar progenitor activity through LRP6 signaling

Research paper thumbnail of Growth of human breast tissues from patient cells in 3D hydrogel scaffolds

Breast cancer research : BCR, 2016

Three-dimensional (3D) cultures have proven invaluable for expanding human tissues for basic rese... more Three-dimensional (3D) cultures have proven invaluable for expanding human tissues for basic research and clinical applications. In both contexts, 3D cultures are most useful when they (1) support the outgrowth of tissues from primary human cells that have not been immortalized through extensive culture or viral infection and (2) include defined, physiologically relevant components. Here we describe a 3D culture system with both of these properties that stimulates the outgrowth of morphologically complex and hormone-responsive mammary tissues from primary human breast epithelial cells. Primary human breast epithelial cells isolated from patient reduction mammoplasty tissues were seeded into 3D hydrogels. The hydrogel scaffolds were composed of extracellular proteins and carbohydrates present in human breast tissue and were cultured in serum-free medium containing only defined components. The physical properties of these hydrogels were determined using atomic force microscopy. Tissue...

Research paper thumbnail of Breast cancer microenvironment and obesity: challenges for therapy

Cancer and Metastasis Reviews

Women with obesity who develop breast cancer have a worsened prognosis with diminished survival r... more Women with obesity who develop breast cancer have a worsened prognosis with diminished survival rates and increased rates of metastasis. Obesity is also associated with decreased breast cancer response to endocrine and chemotherapeutic treatments. Studies utilizing multiple in vivo models of obesity as well as human breast tumors have enhanced our understanding of how obesity alters the breast tumor microenvironment. Changes in the complement and function of adipocytes, adipose-derived stromal cells, immune cells, and endothelial cells and remodeling of the extracellular matrix all contribute to the rapid growth of breast tumors in the context of obesity. Interactions of these cells enhance secretion of cytokines and adipokines as well as local levels of estrogen within the breast tumor microenvironment that promote resistance to multiple therapies. In this review, we will discuss our current understanding of the impact of obesity on the breast tumor microenvironment, how obesity-in...

Research paper thumbnail of Obesity-Activated Lung Stromal Cells Promote Myeloid Lineage Cell Accumulation and Breast Cancer Metastasis

Cancers

Obesity is correlated with increased incidence of breast cancer metastasis; however, the mechanis... more Obesity is correlated with increased incidence of breast cancer metastasis; however, the mechanisms underlying how obesity promotes metastasis are unclear. In a diet-induced obese mouse model, obesity enhanced lung metastasis in both the presence and absence of primary mammary tumors and increased recruitment of myeloid lineage cells into the lungs. In the absence of tumors, obese mice demonstrated increased numbers of myeloid lineage cells and elevated collagen fibers within the lung stroma, reminiscent of premetastatic niches formed by primary tumors. Lung stromal cells isolated from obese tumor-naïve mice showed increased proliferation, contractility, and expression of extracellular matrix, inflammatory markers and transforming growth factor beta-1 (TGFβ1). Conditioned media from lung stromal cells from obese mice promoted myeloid lineage cell migration in vitro in response to colony-stimulating factor 2 (CSF2) expression and enhanced invasion of tumor cells. Together, these resu...