Enhancing the purity and intrinsic properties of ovine dermal papilla cells through flow cytometry sorting and cellular interactions
Objective Dermal Papilla Cells (DPCs) play a crucial role in regulating hair follicle development and serve as a valuable in vitro model for screening and analyzing the genes associated with this process. However, current methods for isolating ovine DPCs primarily rely on mechanical techniques, whic...
Saved in:
| Main Authors: | , , , , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
Asian-Australasian Association of Animal Production Societies
2025-07-01
|
| Series: | Animal Bioscience |
| Subjects: | |
| Online Access: | http://www.animbiosci.org/upload/pdf/ab-24-0805.pdf |
| Tags: |
Add Tag
No Tags, Be the first to tag this record!
|
| _version_ | 1849417044895727616 |
|---|---|
| author | Yuan Gou Tingyan Hu Lijuan Zhu Xiaoyang Lv Yutao Li Rui Su Zhenghai Song Shanhe Wang Wei Sun |
| author_facet | Yuan Gou Tingyan Hu Lijuan Zhu Xiaoyang Lv Yutao Li Rui Su Zhenghai Song Shanhe Wang Wei Sun |
| author_sort | Yuan Gou |
| collection | DOAJ |
| description | Objective Dermal Papilla Cells (DPCs) play a crucial role in regulating hair follicle development and serve as a valuable in vitro model for screening and analyzing the genes associated with this process. However, current methods for isolating ovine DPCs primarily rely on mechanical techniques, which present several limitations. The aim of this study is to establish a method for isolating and culturing ovine DPCs with high purity and retaining their intrinsic properties. Methods We identified sheep DPC membrane-specific genes using single-cell transcriptomic data, validated by immunostaining and flow cytometry. Antibody-labeled DPCs were isolated, cultured, and assessed via fluorescence-activated cell sorting (FACS), comparing their purity with conventional mechanical isolation. Mechanically isolated and flow-sorted DPCs were analyzed through agglutination, cell counting kit (CCK-8), and EDU staining. Furthermore, we examined the biological properties of isolated DPCs in conditioned media using CCK-8, EDU, and quantitative reverse transcription polymerase chain reaction assays. Results PDGFRA was identified as a marker for ovine DPCs. Flow cytometry showed that PDGFRA-labeled DPCs made up 1.54% of the hair follicle cell population, with 1.92% live DPCs obtained via FACS. The isolated DPCs demonstrated agglutination and were positive for ALP, Versican, and α-SMA. Antibody labeling yielded higher DPC purity compared to mechanical isolation, highlighting its efficiency. Accordingly, the addition of conditioned media from mechanically isolated DPCs significantly enhanced agglutination, cell viability, proliferation, and inductive capacity of the sorted DPCs. However, as the number of passages increased, the sorted DPCs demonstrated significant disadvantages in cell agglutination, proliferation rate, and viability compared to mechanically isolated DPCs. Accordingly, the addition of conditioned media from mechanically isolated DPCs significantly enhanced agglutination, cell viability, proliferation, and inductive capacity of the sorted DPCs. Conclusion This study highlights the effectiveness of antibody labeling and flow cytometry for isolating functionally pure DPCs, as well as the potential of conditioned media to maintain the functional properties of these cells. |
| format | Article |
| id | doaj-art-801608ebe23b4be285f5d0452fe3a7e7 |
| institution | Kabale University |
| issn | 2765-0189 2765-0235 |
| language | English |
| publishDate | 2025-07-01 |
| publisher | Asian-Australasian Association of Animal Production Societies |
| record_format | Article |
| series | Animal Bioscience |
| spelling | doaj-art-801608ebe23b4be285f5d0452fe3a7e72025-08-20T03:32:57ZengAsian-Australasian Association of Animal Production SocietiesAnimal Bioscience2765-01892765-02352025-07-013871342135510.5713/ab.24.080525452Enhancing the purity and intrinsic properties of ovine dermal papilla cells through flow cytometry sorting and cellular interactionsYuan Gou0Tingyan Hu1Lijuan Zhu2Xiaoyang Lv3Yutao Li4Rui Su5Zhenghai Song6Shanhe Wang7Wei Sun8 College of Animal Science and Technology, Yangzhou University, Yangzhou, China College of Animal Science and Technology, Yangzhou University, Yangzhou, China College of Animal Science and Technology, Yangzhou University, Yangzhou, China Joint International Research Laboratory of Agriculture and Agri-Product Safety, Ministry of Education of China, Yangzhou University, Yangzhou, China CSIRO Agriculture and Food, Brisbane, Australia Suzhou Sheep Breeding Farm, Suzhou City, Jiangsu, China Dongshan Animal Epidemic Prevention Station, Suzhou, China College of Animal Science and Technology, Yangzhou University, Yangzhou, China College of Animal Science and Technology, Yangzhou University, Yangzhou, ChinaObjective Dermal Papilla Cells (DPCs) play a crucial role in regulating hair follicle development and serve as a valuable in vitro model for screening and analyzing the genes associated with this process. However, current methods for isolating ovine DPCs primarily rely on mechanical techniques, which present several limitations. The aim of this study is to establish a method for isolating and culturing ovine DPCs with high purity and retaining their intrinsic properties. Methods We identified sheep DPC membrane-specific genes using single-cell transcriptomic data, validated by immunostaining and flow cytometry. Antibody-labeled DPCs were isolated, cultured, and assessed via fluorescence-activated cell sorting (FACS), comparing their purity with conventional mechanical isolation. Mechanically isolated and flow-sorted DPCs were analyzed through agglutination, cell counting kit (CCK-8), and EDU staining. Furthermore, we examined the biological properties of isolated DPCs in conditioned media using CCK-8, EDU, and quantitative reverse transcription polymerase chain reaction assays. Results PDGFRA was identified as a marker for ovine DPCs. Flow cytometry showed that PDGFRA-labeled DPCs made up 1.54% of the hair follicle cell population, with 1.92% live DPCs obtained via FACS. The isolated DPCs demonstrated agglutination and were positive for ALP, Versican, and α-SMA. Antibody labeling yielded higher DPC purity compared to mechanical isolation, highlighting its efficiency. Accordingly, the addition of conditioned media from mechanically isolated DPCs significantly enhanced agglutination, cell viability, proliferation, and inductive capacity of the sorted DPCs. However, as the number of passages increased, the sorted DPCs demonstrated significant disadvantages in cell agglutination, proliferation rate, and viability compared to mechanically isolated DPCs. Accordingly, the addition of conditioned media from mechanically isolated DPCs significantly enhanced agglutination, cell viability, proliferation, and inductive capacity of the sorted DPCs. Conclusion This study highlights the effectiveness of antibody labeling and flow cytometry for isolating functionally pure DPCs, as well as the potential of conditioned media to maintain the functional properties of these cells.http://www.animbiosci.org/upload/pdf/ab-24-0805.pdfcellular interactionsdermal papilla cells (dpcs)flow cytometryhair folliclepdgfra |
| spellingShingle | Yuan Gou Tingyan Hu Lijuan Zhu Xiaoyang Lv Yutao Li Rui Su Zhenghai Song Shanhe Wang Wei Sun Enhancing the purity and intrinsic properties of ovine dermal papilla cells through flow cytometry sorting and cellular interactions Animal Bioscience cellular interactions dermal papilla cells (dpcs) flow cytometry hair follicle pdgfra |
| title | Enhancing the purity and intrinsic properties of ovine dermal papilla cells through flow cytometry sorting and cellular interactions |
| title_full | Enhancing the purity and intrinsic properties of ovine dermal papilla cells through flow cytometry sorting and cellular interactions |
| title_fullStr | Enhancing the purity and intrinsic properties of ovine dermal papilla cells through flow cytometry sorting and cellular interactions |
| title_full_unstemmed | Enhancing the purity and intrinsic properties of ovine dermal papilla cells through flow cytometry sorting and cellular interactions |
| title_short | Enhancing the purity and intrinsic properties of ovine dermal papilla cells through flow cytometry sorting and cellular interactions |
| title_sort | enhancing the purity and intrinsic properties of ovine dermal papilla cells through flow cytometry sorting and cellular interactions |
| topic | cellular interactions dermal papilla cells (dpcs) flow cytometry hair follicle pdgfra |
| url | http://www.animbiosci.org/upload/pdf/ab-24-0805.pdf |
| work_keys_str_mv | AT yuangou enhancingthepurityandintrinsicpropertiesofovinedermalpapillacellsthroughflowcytometrysortingandcellularinteractions AT tingyanhu enhancingthepurityandintrinsicpropertiesofovinedermalpapillacellsthroughflowcytometrysortingandcellularinteractions AT lijuanzhu enhancingthepurityandintrinsicpropertiesofovinedermalpapillacellsthroughflowcytometrysortingandcellularinteractions AT xiaoyanglv enhancingthepurityandintrinsicpropertiesofovinedermalpapillacellsthroughflowcytometrysortingandcellularinteractions AT yutaoli enhancingthepurityandintrinsicpropertiesofovinedermalpapillacellsthroughflowcytometrysortingandcellularinteractions AT ruisu enhancingthepurityandintrinsicpropertiesofovinedermalpapillacellsthroughflowcytometrysortingandcellularinteractions AT zhenghaisong enhancingthepurityandintrinsicpropertiesofovinedermalpapillacellsthroughflowcytometrysortingandcellularinteractions AT shanhewang enhancingthepurityandintrinsicpropertiesofovinedermalpapillacellsthroughflowcytometrysortingandcellularinteractions AT weisun enhancingthepurityandintrinsicpropertiesofovinedermalpapillacellsthroughflowcytometrysortingandcellularinteractions |