Isolation and culture of vascular wall-resident cd34+ stem/progenitor cells
Objective: The aim of this study is to observe the maintenance of stem cell properties of purified CD34-positive cells in vessel walls during in vitro expansion. Materials and Methods: Cells that migrated from the adventitial tissues of rat were collected and purified by microbead selection method t...
| Main Authors: | , , , |
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| Format: | Article |
| Language: | English |
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Wolters Kluwer Health/LWW
2019-01-01
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| Series: | Cardiology Plus |
| Subjects: | |
| Online Access: | http://www.cardiologyplus.org/article.asp?issn=2470-7511;year=2019;volume=4;issue=4;spage=116;epage=120;aulast=Wu |
| _version_ | 1811323654113329152 |
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| author | Yan Wu Ruo-Nan Zhang Sen-Zhao Jun-Ming Tang |
| author_facet | Yan Wu Ruo-Nan Zhang Sen-Zhao Jun-Ming Tang |
| author_sort | Yan Wu |
| collection | DOAJ |
| description | Objective: The aim of this study is to observe the maintenance of stem cell properties of purified CD34-positive cells in vessel walls during in vitro expansion. Materials and Methods: Cells that migrated from the adventitial tissues of rat were collected and purified by microbead selection method to obtain CD34+ vascular wall-resident stem (VRS)/progenitor cell (PC). Those purified CD34+ VRS/PCs were evaluated by flow cytometry and immunofluorescent staining. The CD34+ VRS/PCs were continuously cultured until passage P3. Each passaged cell was evaluated by flow cytometry with anti-CD34. Results: After microbead selection, the CD34+ cells reached 88.07% ± 4.36% and these cells expressed neither endothelial (CD31) nor mature smooth muscle cell (smooth muscle-myosin heavy chain and SM22α) markers. Incubation of the purified CD34+ VRS/PCs at a density of 1.5 × 105 cells/100-mm dish, resulted in a gradual reduction of CD34-positive traits when passaged in vitro, starting at P1. Interestingly, the purified primary CD34+ VRS/PCs at a density of 1.0 × 104 cells per 100-mm dish show the traits of colony form growth, and P1 passaged cells were 79.2% ± 2.15% positive for CD34, then gradually lost the traits of CD34-positive cells when passaged in vitro. Conclusions: High purity CD34+ VRS/PCs can be obtained by magnetic bead screening. In vitro, low cell densities contribute to the maintenance of CD34+ VRS/PC traits. |
| first_indexed | 2024-04-13T13:58:59Z |
| format | Article |
| id | doaj.art-c81e29ee134542af90145fe7c27fde00 |
| institution | Directory Open Access Journal |
| issn | 2470-7511 2470-752X |
| language | English |
| last_indexed | 2024-04-13T13:58:59Z |
| publishDate | 2019-01-01 |
| publisher | Wolters Kluwer Health/LWW |
| record_format | Article |
| series | Cardiology Plus |
| spelling | doaj.art-c81e29ee134542af90145fe7c27fde002022-12-22T02:44:06ZengWolters Kluwer Health/LWWCardiology Plus2470-75112470-752X2019-01-014411612010.4103/cp.cp_19_19Isolation and culture of vascular wall-resident cd34+ stem/progenitor cellsYan WuRuo-Nan ZhangSen-ZhaoJun-Ming TangObjective: The aim of this study is to observe the maintenance of stem cell properties of purified CD34-positive cells in vessel walls during in vitro expansion. Materials and Methods: Cells that migrated from the adventitial tissues of rat were collected and purified by microbead selection method to obtain CD34+ vascular wall-resident stem (VRS)/progenitor cell (PC). Those purified CD34+ VRS/PCs were evaluated by flow cytometry and immunofluorescent staining. The CD34+ VRS/PCs were continuously cultured until passage P3. Each passaged cell was evaluated by flow cytometry with anti-CD34. Results: After microbead selection, the CD34+ cells reached 88.07% ± 4.36% and these cells expressed neither endothelial (CD31) nor mature smooth muscle cell (smooth muscle-myosin heavy chain and SM22α) markers. Incubation of the purified CD34+ VRS/PCs at a density of 1.5 × 105 cells/100-mm dish, resulted in a gradual reduction of CD34-positive traits when passaged in vitro, starting at P1. Interestingly, the purified primary CD34+ VRS/PCs at a density of 1.0 × 104 cells per 100-mm dish show the traits of colony form growth, and P1 passaged cells were 79.2% ± 2.15% positive for CD34, then gradually lost the traits of CD34-positive cells when passaged in vitro. Conclusions: High purity CD34+ VRS/PCs can be obtained by magnetic bead screening. In vitro, low cell densities contribute to the maintenance of CD34+ VRS/PC traits.http://www.cardiologyplus.org/article.asp?issn=2470-7511;year=2019;volume=4;issue=4;spage=116;epage=120;aulast=Wucd34+ vascular wall-resident stem/progenitor cellsculturemagnetic bead screening |
| spellingShingle | Yan Wu Ruo-Nan Zhang Sen-Zhao Jun-Ming Tang Isolation and culture of vascular wall-resident cd34+ stem/progenitor cells Cardiology Plus cd34+ vascular wall-resident stem/progenitor cells culture magnetic bead screening |
| title | Isolation and culture of vascular wall-resident cd34+ stem/progenitor cells |
| title_full | Isolation and culture of vascular wall-resident cd34+ stem/progenitor cells |
| title_fullStr | Isolation and culture of vascular wall-resident cd34+ stem/progenitor cells |
| title_full_unstemmed | Isolation and culture of vascular wall-resident cd34+ stem/progenitor cells |
| title_short | Isolation and culture of vascular wall-resident cd34+ stem/progenitor cells |
| title_sort | isolation and culture of vascular wall resident cd34 stem progenitor cells |
| topic | cd34+ vascular wall-resident stem/progenitor cells culture magnetic bead screening |
| url | http://www.cardiologyplus.org/article.asp?issn=2470-7511;year=2019;volume=4;issue=4;spage=116;epage=120;aulast=Wu |
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