3D culture model and computer-assisted videomicroscopy to analyze migratory behavior of noninvasive and invasive bronchial epithelial cells

Abstract

To date, most of the studies in the field of cell migration have been applied to two-dimensional (2D) models. To mimic the three-dimensional (3D) conditions similar to those observed in vivo during tumor invasion, we developed a 3D model of cell migration in which cells were embedded in a collagen I matrix placed in a double-compartment chamber. Using time-lapse videomicroscopy and interactive cell tracking in a four-dimensional data set, we determined the cell trajectories and their migration kinetics. We compared the 2D and 3D migratory behavior of a noninvasive cell line (16HBE) with the migratory behavior of an invasive cell line (BZR). Our results show that the 3D migration kinetics of the noninvasive cell line were lower than the migration kinetics of the invasive cell line. In contrast, in 2D models, no significant difference was observed between the two cell lines. To validate our 3D model, we further investigated the effect of epidermal growth factor (EGF), a promoter of tumor cell motility and invasion on the noninvasive cell line (16HBE). EGF increased significantly the migration kinetics of the noninvasive cell line. Our results show that the 3D model of cell migration allowed us to differentiate the migratory behavior of invasive and noninvasive cells and that such a model can help in the development of molecular targeted therapy as it approaches the in vivo conditions.

Invasion of carcinoma cells into reconstituted type I collagen gels: Visual real-time analysis by time-lapse microscopy

“…In conclusion, the present method utilizing timelapse microscopy is less involved and simpler than the previous methods (14,16), thus providing a useful system for further evaluating major components of ESM, growth factors, cytokines, and other possible microenvironment factors. Preliminary results on effects of ESM components and insulin-like growth factor-I on MCF-7 invasion have been previously presented at 12th International Congress of Endocrinology in 2004 (18).…”

“…Earlier studies using fibroblasts suggested the possible implication of this assay for cancer cell invasion (8,11). After rapid advances in cancer biology, several 3-dimentional culture systems have been reported for studying cancer cell invasion (12)(13)(14)(15)(16). Of those, the real-time assay system under basal conditions described in this report is the most basic and simplest in vitro model for effectively assessing the invasive behaviors of individual cancer cells.…”

Invasion of carcinoma cells into reconstituted type I collagen gels: Visual real-time analysis by time-lapse microscopy

“…In conclusion, the present method utilizing timelapse microscopy is less involved and simpler than the previous methods (14,16), thus providing a useful system for further evaluating major components of ESM, growth factors, cytokines, and other possible microenvironment factors. Preliminary results on effects of ESM components and insulin-like growth factor-I on MCF-7 invasion have been previously presented at 12th International Congress of Endocrinology in 2004 (18).…”

“…Earlier studies using fibroblasts suggested the possible implication of this assay for cancer cell invasion (8,11). After rapid advances in cancer biology, several 3-dimentional culture systems have been reported for studying cancer cell invasion (12)(13)(14)(15)(16). Of those, the real-time assay system under basal conditions described in this report is the most basic and simplest in vitro model for effectively assessing the invasive behaviors of individual cancer cells.…”

A Formative Evaluation of Two Evidence‐Based Psychotherapies for PTSD in VA Residential Treatment Programs

“…Future investigations might examine if the various ways these components are packaged have comparable effects on patient outcomes. This approach may be a better fit for practical, evolving and adaptive clinical settings (Glasgow, & Chambers, 2012). …”

3D collagen type I matrix inhibits the antimigratory effect of doxorubicin

“…Cell migration speed and individual trajectories were determined using time-lapse videomicroscopy and interactive cell tracking in a four-dimensional data set [25]. It is demonstrated that cell migration speed is higher in 3D compared to plastic substrate (25 μm/h versus 18 μm/h, p < 0.01 data not shown).…”

Invasion of carcinoma cells into reconstituted type I collagen gels: Visual real-time analysis by time-lapse microscopy

“…In conclusion, the present method utilizing timelapse microscopy is less involved and simpler than the previous methods (14,16), thus providing a useful system for further evaluating major components of ESM, growth factors, cytokines, and other possible microenvironment factors. Preliminary results on effects of ESM components and insulin-like growth factor-I on MCF-7 invasion have been previously presented at 12th International Congress of Endocrinology in 2004 (18).…”

“…Earlier studies using fibroblasts suggested the possible implication of this assay for cancer cell invasion (8,11). After rapid advances in cancer biology, several 3-dimentional culture systems have been reported for studying cancer cell invasion (12)(13)(14)(15)(16). Of those, the real-time assay system under basal conditions described in this report is the most basic and simplest in vitro model for effectively assessing the invasive behaviors of individual cancer cells.…”

A Formative Evaluation of Two Evidence‐Based Psychotherapies for PTSD in VA Residential Treatment Programs

“…Future investigations might examine if the various ways these components are packaged have comparable effects on patient outcomes. This approach may be a better fit for practical, evolving and adaptive clinical settings (Glasgow, & Chambers, 2012). …”

3D collagen type I matrix inhibits the antimigratory effect of doxorubicin

“…Cell migration speed and individual trajectories were determined using time-lapse videomicroscopy and interactive cell tracking in a four-dimensional data set [25]. It is demonstrated that cell migration speed is higher in 3D compared to plastic substrate (25 μm/h versus 18 μm/h, p < 0.01 data not shown).…”

Invasion of carcinoma cells into reconstituted type I collagen gels: Visual real-time analysis by time-lapse microscopy

“…In conclusion, the present method utilizing timelapse microscopy is less involved and simpler than the previous methods (14,16), thus providing a useful system for further evaluating major components of ESM, growth factors, cytokines, and other possible microenvironment factors. Preliminary results on effects of ESM components and insulin-like growth factor-I on MCF-7 invasion have been previously presented at 12th International Congress of Endocrinology in 2004 (18).…”

“…Earlier studies using fibroblasts suggested the possible implication of this assay for cancer cell invasion (8,11). After rapid advances in cancer biology, several 3-dimentional culture systems have been reported for studying cancer cell invasion (12)(13)(14)(15)(16). Of those, the real-time assay system under basal conditions described in this report is the most basic and simplest in vitro model for effectively assessing the invasive behaviors of individual cancer cells.…”

A Formative Evaluation of Two Evidence‐Based Psychotherapies for PTSD in VA Residential Treatment Programs

“…Future investigations might examine if the various ways these components are packaged have comparable effects on patient outcomes. This approach may be a better fit for practical, evolving and adaptive clinical settings (Glasgow, & Chambers, 2012). …”

3D collagen type I matrix inhibits the antimigratory effect of doxorubicin

“…Cell migration speed and individual trajectories were determined using time-lapse videomicroscopy and interactive cell tracking in a four-dimensional data set [25]. It is demonstrated that cell migration speed is higher in 3D compared to plastic substrate (25 μm/h versus 18 μm/h, p < 0.01 data not shown).…”