The Experts below are selected from a list of 30267 Experts worldwide ranked by ideXlab platform
Stefanie S Jeffrey - One of the best experts on this subject based on the ideXlab platform.
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label free isolation of prostate circulating tumor cells using vortex microfluidic technology
npj Precision Oncology, 2017Co-Authors: Corinne Renier, Edward Pao, James Che, Haiyan E Liu, Clementine A Lemaire, Melissa Matsumoto, Melanie Triboulet, Sandy Srivinas, Stefanie S Jeffrey, Matthew RettigAbstract:There has been increased interest in utilizing non-invasive "liquid biopsies" to identify biomarkers for cancer prognosis and monitoring, and to isolate genetic material that can predict response to targeted therapies. Circulating tumor cells (CTCs) have emerged as such a biomarker providing both genetic and phenotypic information about tumor evolution, potentially from both primary and metastatic sites. Currently, available CTC isolation approaches, including immunoaffinity and size-based filtration, have focused on High Capture Efficiency but with lower purity and often long and manual sample preparation, which limits the use of Captured CTCs for downstream analyses. Here, we describe the use of the microfluidic Vortex Chip for size-based isolation of CTCs from 22 patients with advanced prostate cancer and, from an enumeration study on 18 of these patients, find that we can Capture CTCs with High purity (from 1.74 to 37.59%) and Efficiency (from 1.88 to 93.75 CTCs/7.5 mL) in less than 1 h. Interestingly, more atypical large circulating cells were identified in five age-matched healthy donors (46-77 years old; 1.25-2.50 CTCs/7.5 mL) than in five healthy donors <30 years old (21-27 years old; 0.00 CTC/7.5 mL). Using a threshold calculated from the five age-matched healthy donors (3.37 CTCs/mL), we identified CTCs in 80% of the prostate cancer patients. We also found that a fraction of the cells collected (11.5%) did not express epithelial prostate markers (cytokeratin and/or prostate-specific antigen) and that some instead expressed markers of epithelial-mesenchymal transition, i.e., vimentin and N-cadherin. We also show that the purity and DNA yield of isolated cells is amenable to targeted amplification and next-generation sequencing, without whole genome amplification, identifying unique mutations in 10 of 15 samples and 0 of 4 healthy samples.
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abstract 1525 vortex technology for label free enrichment of ctc from mouse xenograft models
Cancer Research, 2016Co-Authors: Kyra Heirich, Corinne Renier, Melanie Triboulet, Vishnu C Ramani, Elodie Sollier, Stefanie S JeffreyAbstract:Background Non-invasive liquid biopsies, such as CTCs, have been of growing interest due to their potential use in cancer detection, prognosis, and monitoring of therapeutic resistance [1]. Beyond enumeration, characterization of CTCs could help guide treatment selection and the development of targeted cancer therapy. Here, we show that Vortex technology can be used successfully for the size-based Capture of CTCs in a preclinical mouse model of breast cancer. Method To establish that human epithelial cancer cells can be reliably detected in small volumes of mice blood, 50-100 MDA-MB231 and MCF7 breast cancer cells were spiked in 500 μL mice blood, diluted 20 fold, and processed through Vortex chip [2]. Recovered cells were immunostained (CK, CD45, DAPI), and enumerated. For breast cancer xenograft model, 8×106 MDA-MB-231-fLuc/GFP cells were implanted orthotopically into the mammary fat pad of NOD-SCID Gamma mice (n = 35). Tumors were measured in 2 dimensions 3 times/week and tumor volume calculated. Blood from cardiac puncture (500 μl) and lateral saphenous vein (100 μl) was collected starting 1 week post implantation, diluted 40X and processed. Mice were euthanized, organs harvested, formalin fixed, paraffin embedded and HE frequency 1/3), with number and frequency increasing over time up to 147-485 clusters/100 μl by day 42. No CTC were recovered from lateral saphenous vein blood until day 28 post implantation, and their number remained low (mean 2.15±0.65). Microscopic metastases were evident in lung of all mice starting at day 28 and in liver of all mice starting at day 35. Conclusion CTCs were isolated in a label-free manner from mice blood with both High Capture Efficiency and purity. In a preclinical model of metastatic breast cancer, CTC counts correlated well with primary tumor volume and metastases occurrence. Thus the Vortex chip appears to be well suited for the enrichment of CTCs from murine xenograft models. Future works will focus on mice implanted with patient derived xenograft (PDX) and their therapeutic response. Information gathered from these studies should facilitate discovery of new therapeutic targets and the development of personalized medicine. [1] Ignatiadis et al. Clin Cancer Res 2015. [2] Sollier et al. Lab Chip 2014.[K.H. and M.T. contributed equally to this work.] Citation Format: Kyra Heirich, Melanie M. Triboulet, Corinne M. Renier, Vishnu C. Ramani, Elodie Sollier, Stefanie S. Jeffrey. Vortex technology for label-free enrichment of CTC from mouse xenograft models. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 1525.
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abstract 4967 label free collection of prostate circulating tumor cells using microfluidic vortex technology
Cancer Research, 2016Co-Authors: Edward Pao, Corinne Renier, James Che, Clementine A Lemaire, Melanie Triboulet, Sandy Srivinas, Stefanie S Jeffrey, Melissa Matsumoto Di Carlo, Rajan P Kulkarni, Matthew RettigAbstract:BACKGROUND Prostate cancer is among the most common cancers in men worldwide. Better markers than Prostate Specific Antigen (PSA) are still needed for the detection and monitoring of disease progression. Circulating Tumor Cells (CTCs) are shed into the blood stream from primary tumor(s) and may play key roles in the metastatic process. Liquid biopsies have emerged as a promising approach, with a correlation between the CTC numbers and patient prognosis for prostate cancer. CTCs have also been shown to enable early detection of recurrence, and could be potential candidates for guiding cancer therapy in real-time [1]. Current CTC enrichment technologies, including immuno-affinity and size-based filtration methods, have focused on High Capture Efficiency with sometimes tedious sample preparation and overall low purity. METHOD Here, we describe the use of the microfluidic Vortex Chip [2] for rapid and size-based isolation of CTCs from the blood of 23 patients with advanced prostate cancer, and 10 healthy donors; 5 being RESULTS Preliminary work with LNCaP prostate cancer cells spiked in blood showed a 29% Capture Efficiency and 50% purity. In vitro cell assays confirmed that cells enriched with Vortex chip were alive and proliferating for up to 7 days. For 23 patient samples, CTCs were Captured (0.5 - 20 CTCs/mL) with High purity (3.6 - 72.3%), in less than 1H, without prior sample preparation. 11.5% of the cells collected were CK and PSA-negative, but some were identified as undergoing epithelial-mesenchymal transition (EMT) following staining for vimentin and N-cadherin. Few atypical cells were also isolated from age-matched healthy donors (0.7 - 2.8 CTCs/mL), while none was detected in younger healthy donors. Using a threshold calculated from the age-matched healthy donors (3.31 CTCs/mL = mean + 2CV), 70% of the patients were characterized as “positive for CTCs”. No correlation was found between CTC counts and elevated PSA level. CONCLUSION These results demonstrate the ability to rapidly collect pure populations of CTCs in metastatic prostate cancer, independent of surface marker expression, without prior sample preparation. Future studies will use chips with optimized Capture performance, sample recycling, and will include CTC molecular analysis by targeted panel sequencing. A larger cohort of healthy donors is also being examined to determine a statistically-robust CTC baseline for this size-based Capture approach. [1] Scher Hi, et al., J. Clin. Oncol. 2015 [2] Sollier E, et al., Lab Chip 2014 Citation Format: Edward Pao, Corinne Renier, Clementine Lemaire, James Che, Melissa Matsumoto Di Carlo, Melanie Triboulet, Sandy Srivinas, Stefanie S. Jeffrey, Rajan P. Kulkarni, Matthew Rettig, Elodie Sollier, Dino Di Carlo. Label-free collection of prostate circulating tumor cells using microfluidic Vortex technology. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 4967.
Matthew Rettig - One of the best experts on this subject based on the ideXlab platform.
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label free isolation of prostate circulating tumor cells using vortex microfluidic technology
npj Precision Oncology, 2017Co-Authors: Corinne Renier, Edward Pao, James Che, Haiyan E Liu, Clementine A Lemaire, Melissa Matsumoto, Melanie Triboulet, Sandy Srivinas, Stefanie S Jeffrey, Matthew RettigAbstract:There has been increased interest in utilizing non-invasive "liquid biopsies" to identify biomarkers for cancer prognosis and monitoring, and to isolate genetic material that can predict response to targeted therapies. Circulating tumor cells (CTCs) have emerged as such a biomarker providing both genetic and phenotypic information about tumor evolution, potentially from both primary and metastatic sites. Currently, available CTC isolation approaches, including immunoaffinity and size-based filtration, have focused on High Capture Efficiency but with lower purity and often long and manual sample preparation, which limits the use of Captured CTCs for downstream analyses. Here, we describe the use of the microfluidic Vortex Chip for size-based isolation of CTCs from 22 patients with advanced prostate cancer and, from an enumeration study on 18 of these patients, find that we can Capture CTCs with High purity (from 1.74 to 37.59%) and Efficiency (from 1.88 to 93.75 CTCs/7.5 mL) in less than 1 h. Interestingly, more atypical large circulating cells were identified in five age-matched healthy donors (46-77 years old; 1.25-2.50 CTCs/7.5 mL) than in five healthy donors <30 years old (21-27 years old; 0.00 CTC/7.5 mL). Using a threshold calculated from the five age-matched healthy donors (3.37 CTCs/mL), we identified CTCs in 80% of the prostate cancer patients. We also found that a fraction of the cells collected (11.5%) did not express epithelial prostate markers (cytokeratin and/or prostate-specific antigen) and that some instead expressed markers of epithelial-mesenchymal transition, i.e., vimentin and N-cadherin. We also show that the purity and DNA yield of isolated cells is amenable to targeted amplification and next-generation sequencing, without whole genome amplification, identifying unique mutations in 10 of 15 samples and 0 of 4 healthy samples.
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abstract 4967 label free collection of prostate circulating tumor cells using microfluidic vortex technology
Cancer Research, 2016Co-Authors: Edward Pao, Corinne Renier, James Che, Clementine A Lemaire, Melanie Triboulet, Sandy Srivinas, Stefanie S Jeffrey, Melissa Matsumoto Di Carlo, Rajan P Kulkarni, Matthew RettigAbstract:BACKGROUND Prostate cancer is among the most common cancers in men worldwide. Better markers than Prostate Specific Antigen (PSA) are still needed for the detection and monitoring of disease progression. Circulating Tumor Cells (CTCs) are shed into the blood stream from primary tumor(s) and may play key roles in the metastatic process. Liquid biopsies have emerged as a promising approach, with a correlation between the CTC numbers and patient prognosis for prostate cancer. CTCs have also been shown to enable early detection of recurrence, and could be potential candidates for guiding cancer therapy in real-time [1]. Current CTC enrichment technologies, including immuno-affinity and size-based filtration methods, have focused on High Capture Efficiency with sometimes tedious sample preparation and overall low purity. METHOD Here, we describe the use of the microfluidic Vortex Chip [2] for rapid and size-based isolation of CTCs from the blood of 23 patients with advanced prostate cancer, and 10 healthy donors; 5 being RESULTS Preliminary work with LNCaP prostate cancer cells spiked in blood showed a 29% Capture Efficiency and 50% purity. In vitro cell assays confirmed that cells enriched with Vortex chip were alive and proliferating for up to 7 days. For 23 patient samples, CTCs were Captured (0.5 - 20 CTCs/mL) with High purity (3.6 - 72.3%), in less than 1H, without prior sample preparation. 11.5% of the cells collected were CK and PSA-negative, but some were identified as undergoing epithelial-mesenchymal transition (EMT) following staining for vimentin and N-cadherin. Few atypical cells were also isolated from age-matched healthy donors (0.7 - 2.8 CTCs/mL), while none was detected in younger healthy donors. Using a threshold calculated from the age-matched healthy donors (3.31 CTCs/mL = mean + 2CV), 70% of the patients were characterized as “positive for CTCs”. No correlation was found between CTC counts and elevated PSA level. CONCLUSION These results demonstrate the ability to rapidly collect pure populations of CTCs in metastatic prostate cancer, independent of surface marker expression, without prior sample preparation. Future studies will use chips with optimized Capture performance, sample recycling, and will include CTC molecular analysis by targeted panel sequencing. A larger cohort of healthy donors is also being examined to determine a statistically-robust CTC baseline for this size-based Capture approach. [1] Scher Hi, et al., J. Clin. Oncol. 2015 [2] Sollier E, et al., Lab Chip 2014 Citation Format: Edward Pao, Corinne Renier, Clementine Lemaire, James Che, Melissa Matsumoto Di Carlo, Melanie Triboulet, Sandy Srivinas, Stefanie S. Jeffrey, Rajan P. Kulkarni, Matthew Rettig, Elodie Sollier, Dino Di Carlo. Label-free collection of prostate circulating tumor cells using microfluidic Vortex technology. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 4967.
Corinne Renier - One of the best experts on this subject based on the ideXlab platform.
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label free isolation of prostate circulating tumor cells using vortex microfluidic technology
npj Precision Oncology, 2017Co-Authors: Corinne Renier, Edward Pao, James Che, Haiyan E Liu, Clementine A Lemaire, Melissa Matsumoto, Melanie Triboulet, Sandy Srivinas, Stefanie S Jeffrey, Matthew RettigAbstract:There has been increased interest in utilizing non-invasive "liquid biopsies" to identify biomarkers for cancer prognosis and monitoring, and to isolate genetic material that can predict response to targeted therapies. Circulating tumor cells (CTCs) have emerged as such a biomarker providing both genetic and phenotypic information about tumor evolution, potentially from both primary and metastatic sites. Currently, available CTC isolation approaches, including immunoaffinity and size-based filtration, have focused on High Capture Efficiency but with lower purity and often long and manual sample preparation, which limits the use of Captured CTCs for downstream analyses. Here, we describe the use of the microfluidic Vortex Chip for size-based isolation of CTCs from 22 patients with advanced prostate cancer and, from an enumeration study on 18 of these patients, find that we can Capture CTCs with High purity (from 1.74 to 37.59%) and Efficiency (from 1.88 to 93.75 CTCs/7.5 mL) in less than 1 h. Interestingly, more atypical large circulating cells were identified in five age-matched healthy donors (46-77 years old; 1.25-2.50 CTCs/7.5 mL) than in five healthy donors <30 years old (21-27 years old; 0.00 CTC/7.5 mL). Using a threshold calculated from the five age-matched healthy donors (3.37 CTCs/mL), we identified CTCs in 80% of the prostate cancer patients. We also found that a fraction of the cells collected (11.5%) did not express epithelial prostate markers (cytokeratin and/or prostate-specific antigen) and that some instead expressed markers of epithelial-mesenchymal transition, i.e., vimentin and N-cadherin. We also show that the purity and DNA yield of isolated cells is amenable to targeted amplification and next-generation sequencing, without whole genome amplification, identifying unique mutations in 10 of 15 samples and 0 of 4 healthy samples.
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abstract 1525 vortex technology for label free enrichment of ctc from mouse xenograft models
Cancer Research, 2016Co-Authors: Kyra Heirich, Corinne Renier, Melanie Triboulet, Vishnu C Ramani, Elodie Sollier, Stefanie S JeffreyAbstract:Background Non-invasive liquid biopsies, such as CTCs, have been of growing interest due to their potential use in cancer detection, prognosis, and monitoring of therapeutic resistance [1]. Beyond enumeration, characterization of CTCs could help guide treatment selection and the development of targeted cancer therapy. Here, we show that Vortex technology can be used successfully for the size-based Capture of CTCs in a preclinical mouse model of breast cancer. Method To establish that human epithelial cancer cells can be reliably detected in small volumes of mice blood, 50-100 MDA-MB231 and MCF7 breast cancer cells were spiked in 500 μL mice blood, diluted 20 fold, and processed through Vortex chip [2]. Recovered cells were immunostained (CK, CD45, DAPI), and enumerated. For breast cancer xenograft model, 8×106 MDA-MB-231-fLuc/GFP cells were implanted orthotopically into the mammary fat pad of NOD-SCID Gamma mice (n = 35). Tumors were measured in 2 dimensions 3 times/week and tumor volume calculated. Blood from cardiac puncture (500 μl) and lateral saphenous vein (100 μl) was collected starting 1 week post implantation, diluted 40X and processed. Mice were euthanized, organs harvested, formalin fixed, paraffin embedded and HE frequency 1/3), with number and frequency increasing over time up to 147-485 clusters/100 μl by day 42. No CTC were recovered from lateral saphenous vein blood until day 28 post implantation, and their number remained low (mean 2.15±0.65). Microscopic metastases were evident in lung of all mice starting at day 28 and in liver of all mice starting at day 35. Conclusion CTCs were isolated in a label-free manner from mice blood with both High Capture Efficiency and purity. In a preclinical model of metastatic breast cancer, CTC counts correlated well with primary tumor volume and metastases occurrence. Thus the Vortex chip appears to be well suited for the enrichment of CTCs from murine xenograft models. Future works will focus on mice implanted with patient derived xenograft (PDX) and their therapeutic response. Information gathered from these studies should facilitate discovery of new therapeutic targets and the development of personalized medicine. [1] Ignatiadis et al. Clin Cancer Res 2015. [2] Sollier et al. Lab Chip 2014.[K.H. and M.T. contributed equally to this work.] Citation Format: Kyra Heirich, Melanie M. Triboulet, Corinne M. Renier, Vishnu C. Ramani, Elodie Sollier, Stefanie S. Jeffrey. Vortex technology for label-free enrichment of CTC from mouse xenograft models. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 1525.
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abstract 4967 label free collection of prostate circulating tumor cells using microfluidic vortex technology
Cancer Research, 2016Co-Authors: Edward Pao, Corinne Renier, James Che, Clementine A Lemaire, Melanie Triboulet, Sandy Srivinas, Stefanie S Jeffrey, Melissa Matsumoto Di Carlo, Rajan P Kulkarni, Matthew RettigAbstract:BACKGROUND Prostate cancer is among the most common cancers in men worldwide. Better markers than Prostate Specific Antigen (PSA) are still needed for the detection and monitoring of disease progression. Circulating Tumor Cells (CTCs) are shed into the blood stream from primary tumor(s) and may play key roles in the metastatic process. Liquid biopsies have emerged as a promising approach, with a correlation between the CTC numbers and patient prognosis for prostate cancer. CTCs have also been shown to enable early detection of recurrence, and could be potential candidates for guiding cancer therapy in real-time [1]. Current CTC enrichment technologies, including immuno-affinity and size-based filtration methods, have focused on High Capture Efficiency with sometimes tedious sample preparation and overall low purity. METHOD Here, we describe the use of the microfluidic Vortex Chip [2] for rapid and size-based isolation of CTCs from the blood of 23 patients with advanced prostate cancer, and 10 healthy donors; 5 being RESULTS Preliminary work with LNCaP prostate cancer cells spiked in blood showed a 29% Capture Efficiency and 50% purity. In vitro cell assays confirmed that cells enriched with Vortex chip were alive and proliferating for up to 7 days. For 23 patient samples, CTCs were Captured (0.5 - 20 CTCs/mL) with High purity (3.6 - 72.3%), in less than 1H, without prior sample preparation. 11.5% of the cells collected were CK and PSA-negative, but some were identified as undergoing epithelial-mesenchymal transition (EMT) following staining for vimentin and N-cadherin. Few atypical cells were also isolated from age-matched healthy donors (0.7 - 2.8 CTCs/mL), while none was detected in younger healthy donors. Using a threshold calculated from the age-matched healthy donors (3.31 CTCs/mL = mean + 2CV), 70% of the patients were characterized as “positive for CTCs”. No correlation was found between CTC counts and elevated PSA level. CONCLUSION These results demonstrate the ability to rapidly collect pure populations of CTCs in metastatic prostate cancer, independent of surface marker expression, without prior sample preparation. Future studies will use chips with optimized Capture performance, sample recycling, and will include CTC molecular analysis by targeted panel sequencing. A larger cohort of healthy donors is also being examined to determine a statistically-robust CTC baseline for this size-based Capture approach. [1] Scher Hi, et al., J. Clin. Oncol. 2015 [2] Sollier E, et al., Lab Chip 2014 Citation Format: Edward Pao, Corinne Renier, Clementine Lemaire, James Che, Melissa Matsumoto Di Carlo, Melanie Triboulet, Sandy Srivinas, Stefanie S. Jeffrey, Rajan P. Kulkarni, Matthew Rettig, Elodie Sollier, Dino Di Carlo. Label-free collection of prostate circulating tumor cells using microfluidic Vortex technology. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 4967.
Melanie Triboulet - One of the best experts on this subject based on the ideXlab platform.
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label free isolation of prostate circulating tumor cells using vortex microfluidic technology
npj Precision Oncology, 2017Co-Authors: Corinne Renier, Edward Pao, James Che, Haiyan E Liu, Clementine A Lemaire, Melissa Matsumoto, Melanie Triboulet, Sandy Srivinas, Stefanie S Jeffrey, Matthew RettigAbstract:There has been increased interest in utilizing non-invasive "liquid biopsies" to identify biomarkers for cancer prognosis and monitoring, and to isolate genetic material that can predict response to targeted therapies. Circulating tumor cells (CTCs) have emerged as such a biomarker providing both genetic and phenotypic information about tumor evolution, potentially from both primary and metastatic sites. Currently, available CTC isolation approaches, including immunoaffinity and size-based filtration, have focused on High Capture Efficiency but with lower purity and often long and manual sample preparation, which limits the use of Captured CTCs for downstream analyses. Here, we describe the use of the microfluidic Vortex Chip for size-based isolation of CTCs from 22 patients with advanced prostate cancer and, from an enumeration study on 18 of these patients, find that we can Capture CTCs with High purity (from 1.74 to 37.59%) and Efficiency (from 1.88 to 93.75 CTCs/7.5 mL) in less than 1 h. Interestingly, more atypical large circulating cells were identified in five age-matched healthy donors (46-77 years old; 1.25-2.50 CTCs/7.5 mL) than in five healthy donors <30 years old (21-27 years old; 0.00 CTC/7.5 mL). Using a threshold calculated from the five age-matched healthy donors (3.37 CTCs/mL), we identified CTCs in 80% of the prostate cancer patients. We also found that a fraction of the cells collected (11.5%) did not express epithelial prostate markers (cytokeratin and/or prostate-specific antigen) and that some instead expressed markers of epithelial-mesenchymal transition, i.e., vimentin and N-cadherin. We also show that the purity and DNA yield of isolated cells is amenable to targeted amplification and next-generation sequencing, without whole genome amplification, identifying unique mutations in 10 of 15 samples and 0 of 4 healthy samples.
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abstract 1525 vortex technology for label free enrichment of ctc from mouse xenograft models
Cancer Research, 2016Co-Authors: Kyra Heirich, Corinne Renier, Melanie Triboulet, Vishnu C Ramani, Elodie Sollier, Stefanie S JeffreyAbstract:Background Non-invasive liquid biopsies, such as CTCs, have been of growing interest due to their potential use in cancer detection, prognosis, and monitoring of therapeutic resistance [1]. Beyond enumeration, characterization of CTCs could help guide treatment selection and the development of targeted cancer therapy. Here, we show that Vortex technology can be used successfully for the size-based Capture of CTCs in a preclinical mouse model of breast cancer. Method To establish that human epithelial cancer cells can be reliably detected in small volumes of mice blood, 50-100 MDA-MB231 and MCF7 breast cancer cells were spiked in 500 μL mice blood, diluted 20 fold, and processed through Vortex chip [2]. Recovered cells were immunostained (CK, CD45, DAPI), and enumerated. For breast cancer xenograft model, 8×106 MDA-MB-231-fLuc/GFP cells were implanted orthotopically into the mammary fat pad of NOD-SCID Gamma mice (n = 35). Tumors were measured in 2 dimensions 3 times/week and tumor volume calculated. Blood from cardiac puncture (500 μl) and lateral saphenous vein (100 μl) was collected starting 1 week post implantation, diluted 40X and processed. Mice were euthanized, organs harvested, formalin fixed, paraffin embedded and HE frequency 1/3), with number and frequency increasing over time up to 147-485 clusters/100 μl by day 42. No CTC were recovered from lateral saphenous vein blood until day 28 post implantation, and their number remained low (mean 2.15±0.65). Microscopic metastases were evident in lung of all mice starting at day 28 and in liver of all mice starting at day 35. Conclusion CTCs were isolated in a label-free manner from mice blood with both High Capture Efficiency and purity. In a preclinical model of metastatic breast cancer, CTC counts correlated well with primary tumor volume and metastases occurrence. Thus the Vortex chip appears to be well suited for the enrichment of CTCs from murine xenograft models. Future works will focus on mice implanted with patient derived xenograft (PDX) and their therapeutic response. Information gathered from these studies should facilitate discovery of new therapeutic targets and the development of personalized medicine. [1] Ignatiadis et al. Clin Cancer Res 2015. [2] Sollier et al. Lab Chip 2014.[K.H. and M.T. contributed equally to this work.] Citation Format: Kyra Heirich, Melanie M. Triboulet, Corinne M. Renier, Vishnu C. Ramani, Elodie Sollier, Stefanie S. Jeffrey. Vortex technology for label-free enrichment of CTC from mouse xenograft models. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 1525.
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abstract 4967 label free collection of prostate circulating tumor cells using microfluidic vortex technology
Cancer Research, 2016Co-Authors: Edward Pao, Corinne Renier, James Che, Clementine A Lemaire, Melanie Triboulet, Sandy Srivinas, Stefanie S Jeffrey, Melissa Matsumoto Di Carlo, Rajan P Kulkarni, Matthew RettigAbstract:BACKGROUND Prostate cancer is among the most common cancers in men worldwide. Better markers than Prostate Specific Antigen (PSA) are still needed for the detection and monitoring of disease progression. Circulating Tumor Cells (CTCs) are shed into the blood stream from primary tumor(s) and may play key roles in the metastatic process. Liquid biopsies have emerged as a promising approach, with a correlation between the CTC numbers and patient prognosis for prostate cancer. CTCs have also been shown to enable early detection of recurrence, and could be potential candidates for guiding cancer therapy in real-time [1]. Current CTC enrichment technologies, including immuno-affinity and size-based filtration methods, have focused on High Capture Efficiency with sometimes tedious sample preparation and overall low purity. METHOD Here, we describe the use of the microfluidic Vortex Chip [2] for rapid and size-based isolation of CTCs from the blood of 23 patients with advanced prostate cancer, and 10 healthy donors; 5 being RESULTS Preliminary work with LNCaP prostate cancer cells spiked in blood showed a 29% Capture Efficiency and 50% purity. In vitro cell assays confirmed that cells enriched with Vortex chip were alive and proliferating for up to 7 days. For 23 patient samples, CTCs were Captured (0.5 - 20 CTCs/mL) with High purity (3.6 - 72.3%), in less than 1H, without prior sample preparation. 11.5% of the cells collected were CK and PSA-negative, but some were identified as undergoing epithelial-mesenchymal transition (EMT) following staining for vimentin and N-cadherin. Few atypical cells were also isolated from age-matched healthy donors (0.7 - 2.8 CTCs/mL), while none was detected in younger healthy donors. Using a threshold calculated from the age-matched healthy donors (3.31 CTCs/mL = mean + 2CV), 70% of the patients were characterized as “positive for CTCs”. No correlation was found between CTC counts and elevated PSA level. CONCLUSION These results demonstrate the ability to rapidly collect pure populations of CTCs in metastatic prostate cancer, independent of surface marker expression, without prior sample preparation. Future studies will use chips with optimized Capture performance, sample recycling, and will include CTC molecular analysis by targeted panel sequencing. A larger cohort of healthy donors is also being examined to determine a statistically-robust CTC baseline for this size-based Capture approach. [1] Scher Hi, et al., J. Clin. Oncol. 2015 [2] Sollier E, et al., Lab Chip 2014 Citation Format: Edward Pao, Corinne Renier, Clementine Lemaire, James Che, Melissa Matsumoto Di Carlo, Melanie Triboulet, Sandy Srivinas, Stefanie S. Jeffrey, Rajan P. Kulkarni, Matthew Rettig, Elodie Sollier, Dino Di Carlo. Label-free collection of prostate circulating tumor cells using microfluidic Vortex technology. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 4967.
Jae Hee Jung - One of the best experts on this subject based on the ideXlab platform.
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washable antimicrobial polyester aluminum air filter with a High Capture Efficiency and low pressure drop
Journal of Hazardous Materials, 2018Co-Authors: Dong Yun Choi, Ki Joon Heo, Juhee Kang, Sooho Jung, Byung Uk Lee, Hye Moon Lee, Jae Hee JungAbstract:Here, we introduce a reusable bifunctional polyester/aluminum (PET/Al) air filter for the High Efficiency simultaneous Capture and inactivation of airborne microorganisms. Both bacteria of Escherichia coli and Staphylococcus epidermidis were collected on the PET/Al filter with a High Efficiency rate (∼99.99%) via the electrostatic interactions between the charged bacteria and fibers without sacrificing pressure drop. The PET/Al filter experienced a pressure drop approximately 10 times lower per thickness compared with a commercial High-Efficiency particulate air filter. As the Al nanograins grew on the fibers, the antimicrobial activity against airborne E. coli and S. epidermidis improved to ∼94.8% and ∼96.9%, respectively, due to the reinforced hydrophobicity and surface roughness of the filter. Moreover, the Capture and antimicrobial performances were stably maintained during a cyclic washing test of the PET/Al filter, indicative of its reusability. The PET/Al filter shows great potential for use in energy-efficient bioaerosol control systems suitable for indoor environments.