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Updated: May 18, 2026
Murine Dermal Lymphatic Endothelial Cell Isolation
Published on: July 21, 2023
Isolation and characterization of endothelial cells from intramuscular hemangioma
Jae Hwan Cho1, Ilkyu Han, Mi Ra Lee
1Department of Orthopaedic Surgery, Seoul National University Hospital, 101 Daehangno, Jongno Gu, Seoul 110-744, Republic of Korea.
Insights
Endothelial cells (ECs) isolated from intramuscular hemangiomas (IMHs) show increased Tie2 expression. This suggests the Tie2-Akt-FOXO1 pathway is crucial in IMH development.
Area of Science:
- Vascular Biology
- Oncology
- Cell Biology
Background:
- Intramuscular hemangiomas (IMHs) are benign vascular tumors originating from endothelial cell (EC) proliferation.
- Understanding the molecular mechanisms driving IMH development is essential for targeted therapies.
Purpose of the Study:
- To isolate and characterize endothelial cells (ECs) from IMHs.
- To investigate the angiogenic phenotype and functional characteristics of IMH-derived ECs.
- To identify potential signaling pathways involved in IMH pathogenesis.
Main Methods:
- Endothelial cell isolation from IMH tissue via enzymatic digestion and purification.
- Reverse transcriptase polymerase chain reaction (RT-PCR) to assess Tie2 and VEGFR1 expression.
- Cell invasion and proliferation assays to evaluate responses to Angiopoietin 1 (Ang1) and Vascular Endothelial Growth Factor (VEGF).
- Western blot analysis to detect downstream signaling molecules like phosphorylated Akt and FOXO1.
Main Results:
- Isolated IMH ECs exhibited characteristic cobblestone morphology and formed capillary-like structures in vitro.
- RT-PCR revealed significantly higher Tie2 and VEGFR1 expression in IMH ECs compared to controls.
- Ang1-induced Tie2 activation and VEGF-induced VEGFR1 activation promoted EC migration and proliferation.
- Western blot confirmed elevated Tie2 expression in IMH samples and detected phosphorylated Akt and FOXO1 exclusively in hemangioma tissues.
Conclusions:
- Endothelial cell isolation from IMH provides a valuable model for further research.
- Increased Tie2 expression, coupled with activation of the Akt-FOXO1 pathway, appears to be a key factor in the pathogenesis of IMH.
- These findings highlight the Tie2 signaling pathway as a potential therapeutic target for IMH.
Background:
Intramuscular hemangiomas (IMHs) are benign vascular tumors of deep soft tissue characterized by endothelial cell (EC) proliferation. The purpose of this study was to isolate ECs from IMH, characterize their angiogenic phenotype and functional characteristics, and search for a possible signaling pathway related to IMH development.
Methods:
EC Isolation from IMH was performed by digestion, filtration, washing, incubation, and purification in sequence. Tie2 expression was compared between ECs from IMH and controls using reverse transcriptase polymerase chain reaction (RT-PCR). Cell invasion and proliferation assays were used to analyze functional responses of ECs to angiopoietin 1 (Ang1) and vascular endothelial growth factor (VEGF). Expression of downstream targets was analyzed using Western blot analysis.
Results:
Isolated ECs showed typical cobblestone appearance under light microscopy and formed capillary-like tubular structures using Matrigel tube-forming assay. RT-PCR of isolated ECs from six patients showed increased expression of Tie2 and VEGF receptor 1 (VEGFR1) compared with control ECs. Tie2 activation by Ang1 compared with VEGFR1 by VEGF resulted in increased EC migration and proliferation. Western blot analysis showed increased Tie2 expression in hemangioma samples compared with normal ECs. Phosphorylated Akt and phosphorylated forkhead box O1 (FOXO1) expression was observed in hemangioma samples only.
Conclusion:
EC isolation from IMH could be a useful tool for further research. These results suggest that increased Tie2 expression, via Akt-FOXO1 pathway activation, may play an important role in IMH pathogenesis.
