Conditioned medium derived from mesenchymal stem cells overexpressing HPV16 E6E7 dramatically improves ischemic limb.
Mesenchymal stem cells (MSCs) have been shown to secrete cytokines and growth factors required for angiogenesis. Previously, we demonstrated that MSCs expressing HPV16 E6E7 mRNA (E6E7-MSCs) increase life span and differentiation potential and maintain without neoplastic transformation. Whether E6E7-MSCs are sources of molecules for enhancing angiogenesis is unknown. We demonstrated that E6E7-MSC-derived conditioned medium (E6E7-CM) enhanced endothelial cell migration and tube formation compared to primary MSC-derived conditioned medium (primary-CM). Moreover, E6E7-MSCs increased AKT activation and enhanced the release of Interleukin-1β (IL-1β) and vascular endothelial growth factor A (VEGFA). Neutralization of E6E7-CM with antibodies against IL-1β or VEGFA abrogated its effect in enhancing endothelial migration and tube formation. Primary-CM, added with IL-1β and VEGFA, enhanced its ability to increase endothelial migration and tube formation. E6E7-CM was shown to increase the ability to improve blood perfusion in a mouse limb ischemia model. Histological analysis revealed that E6E7-CM prohibited muscle loss or fibrosis and increased endothelial cell counts compared to primary-CM. Similarly, the effects of E6E7-CM in improving perfusion in ischemic limb were also contributed by the increase of IL-1β or VEGFA levels. These results suggest that E6E7-MSCs increase the ability to secrete angiogenic factors via AKT activation, and E6E7-CM is abundant in IL-1β and VEGFA levels and thereby increases the ability to improve blood perfusion and prohibit muscle loss or fibrosis in a mouse limb ischemia model.
Institute of Anatomy, Faculty of Medicine, National Yang-Ming University, Taipei 112, Taiwan; Department of Orthopaedics and Traumatology, Taipei Veterans General Hospital, Taipei 112, Taiwan.
Ming-Chau
MC
Chang
Stem Cell Laboratory, Department of Medical Research, Taipei Veterans General Hospital, Taipei 112, Taiwan.
Ching-Hua
CH
Tsao
Institute of Clinical Medicine, National Yang-Ming University, Taipei 112, Taiwan.
Wei-Hua
WH
Huang
Department of Pathology, Taipei Veterans General Hospital, Taipei 112, Taiwan.
Paul
P
Chih-Hsueh Chen
Institute of Clinical Medicine, National Yang-Ming University, Taipei 112, Taiwan; Institute of Pharmacology, National Yang-Ming University, Taipei 112, Taiwan; Stem Cell Laboratory, Department of Medical Research, Taipei Veterans General Hospital, Taipei 112, Taiwan; Department of Orthopaedics and Traumatology, Taipei Veterans General Hospital, Taipei 112, Taiwan; Institute of Biomedical Sciences, Academia Sinica, Taipei 105, Taiwan. Electronic address: hungsc@vghtpe.gov.tw.
Shih-Chieh
SC
Hung
D000818:Animals; D017077:Culture Media, Conditioned / Q000737:chemistry / Q000494:pharmacology*; D004210:Dissection; D005263:Femoral Artery / Q000601:surgery; D015870:Gene Expression; D052162:Human papillomavirus 16 / Q000235:genetics / Q000254:growth & development; D006801:Humans; D053583:Interleukin-1beta / Q000096:biosynthesis / Q000378:metabolism; D007511:Ischemia / Q000188:drug therapy* / Q000473:pathology; D008297:Male; D059630:Mesenchymal Stem Cells / Q000166:cytology / Q000378:metabolism* / Q000821:virology; D051379:Mice; D008807:Mice, Inbred BALB C; D018919:Neovascularization, Physiologic / Q000187:drug effects*; D009856:Oncogene Proteins, Viral / Q000096:biosynthesis / Q000235:genetics* / Q000378:metabolism; D050725:Papillomavirus E7 Proteins / Q000096:biosynthesis / Q000235:genetics* / Q000378:metabolism; D051057:Proto-Oncogene Proteins c-akt / Q000235:genetics / Q000378:metabolism; D012097:Repressor Proteins / Q000096:biosynthesis / Q000235:genetics* / Q000378:metabolism; D013848:Thigh / Q000098:blood supply* / Q000473:pathology; D042461:Vascular Endothelial Growth Factor A / Q000096:biosynthesis / Q000378:metabolism
D017077:Culture Media, Conditioned; C068651:E6 protein, Human papillomavirus type 16; D053583:Interleukin-1beta; D009856:Oncogene Proteins, Viral; D050725:Papillomavirus E7 Proteins; D012097:Repressor Proteins; C467484:VEGFA protein, human; D042461:Vascular Endothelial Growth Factor A; C059731:oncogene protein E7, Human papillomavirus type 16; D051057:Proto-Oncogene Proteins c-akt
Angiogenic factors; Marrow stromal cells; Multipotent stromal cells; PI3K-AKT; Paracrine effects