A method of isolation and culture of microvascular endothelial cells from mouse skin

Tae, Sung; Talavera, Dodanim; Demir, Erhan; Nath, Anjali K.; Sierra-Honigmann, M. Rocı́o

doi:10.1016/j.mvr.2005.08.002

Cited by 23 publications

(21 citation statements)

References 10 publications

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“…Isolation and Identification of DMVECs from db/db and db/ϩ Mice-This was done as described previously (33) and as detailed in supplemental Fig. S1.…”

Section: Methodsmentioning

confidence: 99%

14S,21R-Dihydroxydocosahexaenoic Acid Remedies Impaired Healing and Mesenchymal Stem Cell Functions in Diabetic Wounds

Tian

Lü

Shah

et al. 2011

Journal of Biological Chemistry

104

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Treatment of diabetes-impaired wound healing remains a major unresolved medical challenge. Here, we identified suppressed formation of a novel reparative lipid mediator 14S,21R-dihydroxydocosa-4Z,7Z,10Z,12E,16Z,19Z-hexaenoic acid (14S,21R-diHDHA) in cutaneous wounds of diabetic db/db mice. These results indicate that diabetes impedes the biosynthetic pathways of 14S,21R-diHDHA in skin wounds. Administration of exogenous 14S,21R-diHDHA to wounds in diabetic animals rescued healing and angiogenesis. When db/db mesenchymal stem cells (MSCs) were administered together with 14S,21R-diHDHA to wounds in diabetic animals, they coacted to accelerate wound re-epithelialization, granulation tissue formation, and synergistically improved vascularization. In the pivotal cellular processes of angiogenesis, 14S,21R-diHDHA enhanced VEGF release, vasculature formation, and migration of db/db dermal microvascular endothelial cells (DMVECs), as well as remedied paracrine angiogenic functions of db/db MSCs, including VEGF secretion and the promotion of DMVEC migration and vasculature formation. Our results show that 14S,21R-diHDHA activates the p38 MAPK pathway in wounds, db/db MSCs, and DMVECs. Overall, the impeded formation of 14S,21R-diHDHA described in this study suggests that diabetes could affect the generation of pro-healing lipid mediators in wound healing. By restoring wound healing and MSC functions, 14S,21R-diHDHA is a new lead for the development of better therapeutics used in treating wounds of diabetics.

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“…Isolation and Identification of DMVECs from db/db and db/ϩ Mice-This was done as described previously (33) and as detailed in supplemental Fig. S1.…”

Section: Methodsmentioning

confidence: 99%

14S,21R-Dihydroxydocosahexaenoic Acid Remedies Impaired Healing and Mesenchymal Stem Cell Functions in Diabetic Wounds

Tian

Lü

Shah

et al. 2011

Journal of Biological Chemistry

104

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“…For real-time quantitative PCR, we used primers and internal standards as previously described [26]; primers for human specimens were purchased from Applied Biosystems (GAPDH, Hs99999905_m1; eNOS, Hs00167166_m1; CD31, Hs00169777_m1; VE-cadherin, Hs00174344_m1). Primers for mouse specimens were as follows: eNOS: 5′-AGAGCCTGCAATTACTACCA-3′, 5′-GTGGATTTGCTGCTCTGTAG-3′; GAPDH: 5′-AACGACCCCTTCATTGAC-3′, 5′-TCCACGACATACTCAGCAC-3′; CD31: 5′-CAGGACCACGTGTTAGTGTT-3′, 5′-ACTCCTGATGGGTTCTGACT-3′ [26, 27]. Quantitative PCR was performed using the SYBR Green supermix (BioRad) and amplified by iQ5 Real-Time PCR detection system (45 cycles, 56°C).…”

Section: Methodsmentioning

confidence: 99%

Endothelial Nitric Oxide Synthase Stimulates Aneurysm Growth in Aged Mice

et al. 2008

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Background/Aims: Age-associated changes in endothelial nitric oxide synthase (eNOS) expression have not been definitively linked to the pathophysiology of aortic aneurysms. We examined the role of eNOS in human patients and an age-appropriate mouse model. Methods: eNOS transcripts and immunodetectable protein were assessed by quantitative PCR and immunohistochemistry in human ascending thoracic aneurysms (n = 29) and referent aortae (n = 31). Carotid aneurysms were induced with CaCl₂ in young adult (3 months) and aged (18 months) C57BL/6 and eNOS-knockout (eNOS-KO) mice. Results: eNOS transcripts and protein were reduced in human aneurysms compared with controls, although aortic eNOS expression also decreased with patient age. Aged wild-type mice had significantly larger aneurysm diameter than young adult mice. Aged wild-type mice had reduced eNOS transcripts and protein compared with young adult mice. Aged eNOS-KO mice had smaller aneurysms compared with aged wild-type mice but similar size aneurysms compared with young eNOS-KO and young wild-type mice. Conclusion: eNOS expression is reduced in both aged human and aged mouse endothelium and eNOS expression is linked to aneurysm expansion in aged but not young adult mice. These findings support the relevance of age-associated changes in eNOS expression in clinical aneurysmal disease.

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“…Primary endothelial cells from human tissues have been used before to explore other diseases, such as type 2 diabetes, although the number of reports is limited, and have not been used in the context of PE (13,44,46). The most commonly reported isolation method is by immunomagnetic beads (16), although others have reported endothelial cell isolation by density gradient (6). We chose the positive-selection isolation method using immunobeads and showed high cell purity levels by endothelial cell marker detection.…”

Section: Discussionmentioning

confidence: 99%

Microvascular endothelial cells from preeclamptic women exhibit altered expression of angiogenic and vasopressor factors

Lee

Nevo

2016

American Journal of Physiology-Heart and Circulatory Physiology

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A method of isolation and culture of microvascular endothelial cells from mouse skin

Cited by 23 publications

References 10 publications

14S,21R-Dihydroxydocosahexaenoic Acid Remedies Impaired Healing and Mesenchymal Stem Cell Functions in Diabetic Wounds

14S,21R-Dihydroxydocosahexaenoic Acid Remedies Impaired Healing and Mesenchymal Stem Cell Functions in Diabetic Wounds

Endothelial Nitric Oxide Synthase Stimulates Aneurysm Growth in Aged Mice

Microvascular endothelial cells from preeclamptic women exhibit altered expression of angiogenic and vasopressor factors

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