Laparoscopic Peritoneal Wash Cytology-Derived Primary Human Mesothelial Cells for In Vitro Cell Culture and Simulation of Human Peritoneum
Peritoneal mucosa of mesothelial cells line the abdominal cavity, surround intestinal organs and the female reproductive organs and are responsible for immunological integrity, organ functionality and regeneration. Peritoneal diseases range from inflammation, adhesions, endometriosis, and cancer. Ef...
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| Format: | Article |
| Language: | English |
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MDPI AG
2021-02-01
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| Series: | Biomedicines |
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| Online Access: | https://www.mdpi.com/2227-9059/9/2/176 |
| _version_ | 1797411624698511360 |
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| author | Myriam Holl Lucas Becker Anna-Lena Keller Nora Feuerer Julia Marzi Daniel A. Carvajal Berrio Peter Jakubowski Felix Neis Jan Pauluschke-Fröhlich Sara Y. Brucker Katja Schenke-Layland Bernhard Krämer Martin Weiss |
| author_facet | Myriam Holl Lucas Becker Anna-Lena Keller Nora Feuerer Julia Marzi Daniel A. Carvajal Berrio Peter Jakubowski Felix Neis Jan Pauluschke-Fröhlich Sara Y. Brucker Katja Schenke-Layland Bernhard Krämer Martin Weiss |
| author_sort | Myriam Holl |
| collection | DOAJ |
| description | Peritoneal mucosa of mesothelial cells line the abdominal cavity, surround intestinal organs and the female reproductive organs and are responsible for immunological integrity, organ functionality and regeneration. Peritoneal diseases range from inflammation, adhesions, endometriosis, and cancer. Efficient technologies to isolate and cultivate healthy patient-derived mesothelial cells with maximal purity enable the generation of capable 2D and 3D as well as in vivo-like microfluidic cell culture models to investigate pathomechanisms and treatment strategies. Here, we describe a new and easily reproducible technique for the isolation and culture of primary human mesothelial cells from laparoscopic peritoneal wash cytology. We established a protocol containing multiple washing and centrifugation steps, followed by cell culture at the highest purity and over multiple passages. Isolated peritoneal mesothelial cells were characterized in detail, utilizing brightfield and immunofluorescence microscopy, flow cytometry as well as Raman microspectroscopy and multivariate data analysis. Thereby, cytokeratin expression enabled specific discrimination from primary peritoneal human fibroblasts. Raman microspectroscopy and imaging were used to study morphology and biochemical properties of primary mesothelial cell culture compared to cryo-fixed and cryo-sectioned peritoneal tissue. |
| first_indexed | 2024-03-09T04:48:50Z |
| format | Article |
| id | doaj.art-62482448dbe94301bf329f0f35a6bf35 |
| institution | Directory Open Access Journal |
| issn | 2227-9059 |
| language | English |
| last_indexed | 2024-03-09T04:48:50Z |
| publishDate | 2021-02-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Biomedicines |
| spelling | doaj.art-62482448dbe94301bf329f0f35a6bf352023-12-03T13:12:39ZengMDPI AGBiomedicines2227-90592021-02-019217610.3390/biomedicines9020176Laparoscopic Peritoneal Wash Cytology-Derived Primary Human Mesothelial Cells for In Vitro Cell Culture and Simulation of Human PeritoneumMyriam Holl0Lucas Becker1Anna-Lena Keller2Nora Feuerer3Julia Marzi4Daniel A. Carvajal Berrio5Peter Jakubowski6Felix Neis7Jan Pauluschke-Fröhlich8Sara Y. Brucker9Katja Schenke-Layland10Bernhard Krämer11Martin Weiss12Department of Women’s Health, Eberhard Karls University, 72076 Tübingen, GermanyDepartment of Women’s Health, Eberhard Karls University, 72076 Tübingen, GermanyNMI Natural and Medical Sciences Institute, University of Tübingen, 72770 Reutlingen, GermanyDepartment of Women’s Health, Eberhard Karls University, 72076 Tübingen, GermanyDepartment of Women’s Health, Eberhard Karls University, 72076 Tübingen, GermanyDepartment of Women’s Health, Eberhard Karls University, 72076 Tübingen, GermanyDepartment of Women’s Health, Eberhard Karls University, 72076 Tübingen, GermanyDepartment of Women’s Health, Eberhard Karls University, 72076 Tübingen, GermanyDepartment of Women’s Health, Eberhard Karls University, 72076 Tübingen, GermanyDepartment of Women’s Health, Eberhard Karls University, 72076 Tübingen, GermanyDepartment of Women’s Health, Eberhard Karls University, 72076 Tübingen, GermanyDepartment of Women’s Health, Eberhard Karls University, 72076 Tübingen, GermanyDepartment of Women’s Health, Eberhard Karls University, 72076 Tübingen, GermanyPeritoneal mucosa of mesothelial cells line the abdominal cavity, surround intestinal organs and the female reproductive organs and are responsible for immunological integrity, organ functionality and regeneration. Peritoneal diseases range from inflammation, adhesions, endometriosis, and cancer. Efficient technologies to isolate and cultivate healthy patient-derived mesothelial cells with maximal purity enable the generation of capable 2D and 3D as well as in vivo-like microfluidic cell culture models to investigate pathomechanisms and treatment strategies. Here, we describe a new and easily reproducible technique for the isolation and culture of primary human mesothelial cells from laparoscopic peritoneal wash cytology. We established a protocol containing multiple washing and centrifugation steps, followed by cell culture at the highest purity and over multiple passages. Isolated peritoneal mesothelial cells were characterized in detail, utilizing brightfield and immunofluorescence microscopy, flow cytometry as well as Raman microspectroscopy and multivariate data analysis. Thereby, cytokeratin expression enabled specific discrimination from primary peritoneal human fibroblasts. Raman microspectroscopy and imaging were used to study morphology and biochemical properties of primary mesothelial cell culture compared to cryo-fixed and cryo-sectioned peritoneal tissue.https://www.mdpi.com/2227-9059/9/2/176mesothelial cellsprimary cell culturein vitro cell culture2D/3D cell culture modelhuman peritoneumlaparoscopy |
| spellingShingle | Myriam Holl Lucas Becker Anna-Lena Keller Nora Feuerer Julia Marzi Daniel A. Carvajal Berrio Peter Jakubowski Felix Neis Jan Pauluschke-Fröhlich Sara Y. Brucker Katja Schenke-Layland Bernhard Krämer Martin Weiss Laparoscopic Peritoneal Wash Cytology-Derived Primary Human Mesothelial Cells for In Vitro Cell Culture and Simulation of Human Peritoneum Biomedicines mesothelial cells primary cell culture in vitro cell culture 2D/3D cell culture model human peritoneum laparoscopy |
| title | Laparoscopic Peritoneal Wash Cytology-Derived Primary Human Mesothelial Cells for In Vitro Cell Culture and Simulation of Human Peritoneum |
| title_full | Laparoscopic Peritoneal Wash Cytology-Derived Primary Human Mesothelial Cells for In Vitro Cell Culture and Simulation of Human Peritoneum |
| title_fullStr | Laparoscopic Peritoneal Wash Cytology-Derived Primary Human Mesothelial Cells for In Vitro Cell Culture and Simulation of Human Peritoneum |
| title_full_unstemmed | Laparoscopic Peritoneal Wash Cytology-Derived Primary Human Mesothelial Cells for In Vitro Cell Culture and Simulation of Human Peritoneum |
| title_short | Laparoscopic Peritoneal Wash Cytology-Derived Primary Human Mesothelial Cells for In Vitro Cell Culture and Simulation of Human Peritoneum |
| title_sort | laparoscopic peritoneal wash cytology derived primary human mesothelial cells for in vitro cell culture and simulation of human peritoneum |
| topic | mesothelial cells primary cell culture in vitro cell culture 2D/3D cell culture model human peritoneum laparoscopy |
| url | https://www.mdpi.com/2227-9059/9/2/176 |
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