Stem Cell Res Ther 9:23(2018) 

Delivery of VEGFA in bone marrow stromal cells seeded in copolymer scaffold enhances angiogenesis, but is inadequate for osteogenesis as compared with the dual delivery of VEGFA and BMP2 in a subcutaneous mouse model

Sunita Sharma


摘要:

Well-characterized primary human BMSC were purchased from StemCell Technologies (cat. number: MSC-001 F; Vancouver, British Columbia, Canada) and expanded using a MesenCult Proliferation Kit (Stem Cell Technologies, Part ID 05411) following the standard culture protocol. All cell culture experiments were carried out in a humidified atmosphere at 37 °C and 5% CO2. For validation of the changes in gene expression induced by adenoviral-mediated delivery of VEGFA alone or in combination with BMP2, commercially available human BMSC from two additional donors were used (henceforth referred to as donor 2 and 3 BMSC), as described in the supporting information (Additional file 1).Transduction of BMSC with ad-GFP, ad-VEGFA, and ad-BMP2 viral particles was efficient (80–90%) and the transduced cells demonstrated similar morphology in monoculture (Fig. 1a–e). Compared to the control ad-GFP BMSC, ad-VEGFA and ad-BMP2 + VEGFA BMSC seeded onto scaffolds expressed significantly higher levels of VEGFA mRNA and protein at day 3 and day 14 (Fig. 1f, h). Similarly, significantly higher levels of BMP2 mRNA and secreted BMP2 protein were found in ad-VEGFA + BMP2 BMSC seeded onto scaffolds at day 3 and 14, compared with the control ad-GFP and ad-VEGFA BMSC (Fig. 1 g, i).We thank Siren Hammer Østvold for the technical assistance during the animal experiments and Dr. Joan Bevenius for English language revision of the manuscript.All animal experiments were approved by the Norwegian Animal Research Authority and conducted according to the European Convention for the Protection of Vertebrates used for Scientific Purposes (local approval number 4940). The normal human oral mucosa specimen was collected after informed written patient consent and was approved by the Committee for Medical and Health Research Ethics in West Norway (ref. 2011/1244/REK vest).Electronic supplementary materialThe online version of this article (10.1186/s13287-018-0778-4) contains supplementary material, which is available to authorized users.
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