Journal: Journal of Tissue Engineering and Regenerative Medicine

Article Title: Oral Presentations

doi: 10.1002/term.1931

Figure Lengend Snippet: Figure 1 SEM surface image (a, 5009) of PGS-PCL scaffold containing encapsulated Res; Young’s Modulus (b); FTIR spectra (c); The expres- sion of MMP9 and MMP2 on EAhy926 endothelial cells under hypoxia conditions after 3 days (d).

Article Snippet: Cancer cell lines were then pre-grown in hydrogels for 5–7 days and then re-seeded into starPEG-heparin hydrogels functionalised with RGD, SDF-1, bFGF and VEGF as spheroids with HUVECs and MSC and grown for 14 days as a tri-culture in Endothelial Cell Growth Medium (ECGM; Promocell).

Techniques:

Journal: Journal of Tissue Engineering and Regenerative Medicine

Article Title: Oral Presentations

doi: 10.1002/term.1931

Figure Lengend Snippet: Figure 1 (A) Tube-in-stent valve in (B) open (systolic) and (C) closed (diastolic) configuration. Materials and methods: The tubular constructs were moulded with fibrin and human umbilical vein cells (60 9 106/mL). They were sewn into a stent (Admedes Schuessler GmbH, Pfonzheim, Germany) and conditioned for 3 weeks in bioreactors. Endothelialization with human umbilical vein cells was performed during the last week of stimulation. After harvesting, the valves underwent crimping for 20 minutes to sim- ulate the catheter based-delivery. The tissue was analyzed by immuno- histochemistry, hydroxyproline assay, scanning electron microscopy (SEM). The performance of the valve was determined in terms of effec- tive orifice area and transvalvular pressure drop under aortic condi- tions. Mechanical properties were quantified as burst strength. Results: After dynamic conditioning the valves (n = 3) were fully func- tional with unobstructed opening (systolic phase) and complete closure (diastolic phase). Tissue analysis showed a homogeneous cell distribu- tion throughout the valve0s thickness and deposition of collagen types I and III oriented along the longitudinal direction. Immunohistochemical staining against CD31 and SEM revealed a confluent endothelial cell layer on the surface of the valves. The crimping procedure did not affect the valvular functionality in terms of effective orifice area and

Article Snippet: Cancer cell lines were then pre-grown in hydrogels for 5–7 days and then re-seeded into starPEG-heparin hydrogels functionalised with RGD, SDF-1, bFGF and VEGF as spheroids with HUVECs and MSC and grown for 14 days as a tri-culture in Endothelial Cell Growth Medium (ECGM; Promocell).

Techniques: Construct, Immunohistochemistry, Hydroxyproline Assay, Electron Microscopy, Immunohistochemical staining, Staining

Journal: Journal of Tissue Engineering and Regenerative Medicine

Article Title: Oral Presentations

doi: 10.1002/term.1931

Figure Lengend Snippet: Figure 1 Necessary steps to transform the iLA to a longterm ambulatory lung assist device. To improve the hemo- and biocomaptiblity of the gasexchange mate- rial Polymethlypentene (PMP), we designed a strategy to seed the fibers with human endothelial cells. Therefore a cell adhesion promot- ing biochemical surface with benzophenone modified heparin and suit- able chemical side chains to perform both, peptide-coupling and covalent conjugation to PMP- membranes was developed. In order to provide cell adhesion sites RGD-peptides were coupled. Human dermal endothelial cells (HDMEC) were seeded to the surface and analyzed with FDA/PI. Results: We succeeded in miniaturization of all hardware components as well as in a new lid and inflow design of the gasexchanger to improve blood distribution (Figure 2a). Figure 2b shows FDA stained vital endothelial cells after 48 h on the PMP fibers under static cell cul- ture conditions condition (37°C, 5% CO2).

Article Snippet: Cancer cell lines were then pre-grown in hydrogels for 5–7 days and then re-seeded into starPEG-heparin hydrogels functionalised with RGD, SDF-1, bFGF and VEGF as spheroids with HUVECs and MSC and grown for 14 days as a tri-culture in Endothelial Cell Growth Medium (ECGM; Promocell).

Techniques: Conjugation Assay, Staining

Journal: Journal of Tissue Engineering and Regenerative Medicine

Article Title: Oral Presentations

doi: 10.1002/term.1931

Figure Lengend Snippet: Figure 1 Co-cultures and triple-cultures were stained for the endothelial marker CD31 followed by quantitative assessment of vascular struc- tures influenced by the addition of EPC.

Article Snippet: Cancer cell lines were then pre-grown in hydrogels for 5–7 days and then re-seeded into starPEG-heparin hydrogels functionalised with RGD, SDF-1, bFGF and VEGF as spheroids with HUVECs and MSC and grown for 14 days as a tri-culture in Endothelial Cell Growth Medium (ECGM; Promocell).

Techniques: Staining, Marker

Journal: Journal of Tissue Engineering and Regenerative Medicine

Article Title: Oral Presentations

doi: 10.1002/term.1931

Figure Lengend Snippet: Figure 2 (A) Co-cultures and triple-cultures on PU-HA after van Kossa staining (B) samples were stained for the endothelial marker CD31 (green) and myeloid marker CD11b (red) in 3D. Discussion and conclusions: Our findings demonstrate that endothelial progenitor cells support mature endothelial cells in the vascularization process in co-culture with MSC whereas the impact on bone formation still needs to be further defined. Acknowledgments: The authors want to thank Katrin Lange, Gabriele Nessenius, and Barbara Pavic for the excellent technical support Disclosures: Authors have nothing to disclose. References Shi et al. Eur Cell and Mater. 2014 Jan 25; 27:64–80.

Article Snippet: Cancer cell lines were then pre-grown in hydrogels for 5–7 days and then re-seeded into starPEG-heparin hydrogels functionalised with RGD, SDF-1, bFGF and VEGF as spheroids with HUVECs and MSC and grown for 14 days as a tri-culture in Endothelial Cell Growth Medium (ECGM; Promocell).

Techniques: Staining, Marker, Co-Culture Assay

Journal: Journal of Tissue Engineering and Regenerative Medicine

Article Title: Oral Presentations

doi: 10.1002/term.1931

Figure Lengend Snippet: Figure 1 Phenotypic characterization of cultured BM-derived macro- phages treated with MSC-conditioned medium. Discussion and conclusions: Our results proved the potential of uncom- mitted MSCs to functionallly mobilize host cells into an induced bone regenerative niche in vivo. In particular, we demonstrated a cross-talk between implanted MSCs and host cells, such as macrophages as well as bone marrow-derived endothelial and mesenchymal precursors,

Article Snippet: Cancer cell lines were then pre-grown in hydrogels for 5–7 days and then re-seeded into starPEG-heparin hydrogels functionalised with RGD, SDF-1, bFGF and VEGF as spheroids with HUVECs and MSC and grown for 14 days as a tri-culture in Endothelial Cell Growth Medium (ECGM; Promocell).

Techniques: Cell Culture, Derivative Assay, In Vivo

Journal: Journal of Tissue Engineering and Regenerative Medicine

Article Title: Oral Presentations

doi: 10.1002/term.1931

Figure Lengend Snippet: Figure 1 (A) Young’s Moduli at 20% strain show increased tunable stiff- ness with the addition of increasing TG. (B) Cell viability is conserved in Fibrin-ECM gels at all stiffness with aECM and lower (but still ~80%) in nECM gels. (C) CPCs interact with ECM (fluorescently labeled green, top) and generate significant cellular networks after 3 wks in culture (bottom). (D) PCR demonstrates increased endothelial differentiation in aECM gels that increases with increasing stiffness. (E) Fibrin-ECM gels are able to be injected into hearts (top) and are retained (Evans blue dye labeled, bottom).

Article Snippet: Cancer cell lines were then pre-grown in hydrogels for 5–7 days and then re-seeded into starPEG-heparin hydrogels functionalised with RGD, SDF-1, bFGF and VEGF as spheroids with HUVECs and MSC and grown for 14 days as a tri-culture in Endothelial Cell Growth Medium (ECGM; Promocell).

Techniques: Labeling, Injection

Journal: Clinical Ophthalmology (Auckland, N.Z.)

Article Title: Novel Needle for Intravitreal Injections Does Not Affect Biological Activity of Anti-VEGF Drugs

doi: 10.2147/OPTH.S557508

Figure Lengend Snippet: Passage of ranibizumab through the NDN or SHN did not change its typical characteristics. After incubation of VEGF-A 165 with ( A ) untreated ranibizumab or ( B ) ranibizumab repeatedly passed through a needle, free growth factor was determined by ELISA. Data from independent experiments were pooled and analyzed as described in Material and Methods, and are shown as scatter plots with means ± standard deviations. *p<0.05, **p<0.01, ***p<0.001, ****) p<0.0001; only statistically significant differences are shown. All measured concentrations of free VEGF-A were very low. ( C – F ) Confluent monolayers of retinal endothelial cells (iBREC) were exposed to vehicle (n=6), VEGF-A 165 (n=8), the growth factor plus ( C and F ) untreated (n=8), ( D and F ) SHN-treated (n=12) or ( E and F ) NDN-treated (n=11) ranibizumab for three days and the cell index was measured continuously. Cell index values were normalized and analyzed as described in Material and Methods. Means ± standard deviations are shown in ( C – E ), means only in ( F ). Ranibizumab efficiently prevented VEGF-A 165 -induced decline of cell index values which was not changed by its passage either through a SHN or the NDN.

Article Snippet: Briefly, cells were cultured in Endothelial Cell Growth Medium MV (ECGM; # C22120 , PromoCell, Heidelberg, Germany) until a confluent cell monolayer was reached three days later, indicated by a constantly high cell index (CI ~ 20).

Techniques: Incubation, Enzyme-linked Immunosorbent Assay

Journal: Bioengineering & Translational Medicine

Article Title: Mesenchymal stem cell extracellular vesicle vascularization bioactivity and production yield are responsive to cell culture substrate stiffness

doi: 10.1002/btm2.10743

Figure Lengend Snippet: Substrate stiffness influences BM‐MSC EV production and bioactivity. (a) EV production as quantified by EVs per cell from BM‐MSCs seeded on Sylgard 184 PDMS substrates with different base‐to‐crosslinker ratios. EVs used for this data were from 1 day of collection and isolated and counted separately from the conditioned media from the other 2 days. After media collection, cells were trypsinized and counted ( n = 3). (b) After a scratch was induced, HUVECs were treated with BM‐MSC EVs from the different substrates or growth or basal media, and percent gap closure after 20 h was evaluated via microscopy ( n = 3). (c) HUVECs were resuspended in EV treatments or growth or basal endothelial media, seeded in Matrigel‐coated wells, and tube formation after 3–6 h was quantified by the number of loops that had formed ( n = 3). All values expressed as mean ± SD. All data are representative of at least three independent experiments ( n = 3). Statistical significance was determined by ANOVA; *p < 0.05, **p < 0.01, ***p < 0.001, and ****p < 0.0001.

Article Snippet: To measure in vitro angiogenesis, 48‐well plates were coated with 60 μL of growth factor reduced Matrigel (Corning; 356230) and incubated at 37°C for 30 min. P4 HUVECs were then seeded at 35,000 cells/well with either endothelial growth media (PromoCell; C‐22121) with 1% penicillin–streptomycin (positive control), endothelial basal media (negative control), or endothelial basal media (PromoCell; C‐22221) with 0.1% FBS and 1% penicillin–streptomycin with 5E9 EVs/mL.

Techniques: Isolation, Microscopy

Journal: Bioengineering & Translational Medicine

Article Title: Mesenchymal stem cell extracellular vesicle vascularization bioactivity and production yield are responsive to cell culture substrate stiffness

doi: 10.1002/btm2.10743

Figure Lengend Snippet: Softer 184:527 PDMS substrates improve the angiogenic bioactivity of BM‐MSC EVs. (a) EV production quantified as EV per cell from BM‐MSCs seeded on each substrate made with different ratios of Sylgard 184 and Sylgard 527 ( n = 2). EVs used for this data were from 1 day of collection and isolated and counted separately from the conditioned media from the other 2 days. After media collection, cells were trypsinized and counted. (b) After a scratch was induced, HUVECs were treated with BM‐MSC EVs from the different substrates or growth or basal media, and percent gap closure after 20 h was evaluated via microscopy ( n = 3). (c) HUVECs were resuspended in the different EV treatments or growth or basal endothelial basal media, and tube formation after 3–6 h was quantified by the number of loops that had formed ( n = 3). All values expressed as mean ± SD. Statistical significance was determined by ANOVA; * p < 0.05, ** p < 0.01, and **** p < 0.0001.

Article Snippet: To measure in vitro angiogenesis, 48‐well plates were coated with 60 μL of growth factor reduced Matrigel (Corning; 356230) and incubated at 37°C for 30 min. P4 HUVECs were then seeded at 35,000 cells/well with either endothelial growth media (PromoCell; C‐22121) with 1% penicillin–streptomycin (positive control), endothelial basal media (negative control), or endothelial basal media (PromoCell; C‐22221) with 0.1% FBS and 1% penicillin–streptomycin with 5E9 EVs/mL.

Techniques: Isolation, Microscopy

Journal: Bioengineering & Translational Medicine

Article Title: Mesenchymal stem cell extracellular vesicle vascularization bioactivity and production yield are responsive to cell culture substrate stiffness

doi: 10.1002/btm2.10743

Figure Lengend Snippet: Substrate stiffness influences BM‐MSC EV production and bioactivity. (a) EV production as quantified by EVs per cell from BM‐MSCs seeded on Sylgard 184 PDMS substrates with different base‐to‐crosslinker ratios. EVs used for this data were from 1 day of collection and isolated and counted separately from the conditioned media from the other 2 days. After media collection, cells were trypsinized and counted ( n = 3). (b) After a scratch was induced, HUVECs were treated with BM‐MSC EVs from the different substrates or growth or basal media, and percent gap closure after 20 h was evaluated via microscopy ( n = 3). (c) HUVECs were resuspended in EV treatments or growth or basal endothelial media, seeded in Matrigel‐coated wells, and tube formation after 3–6 h was quantified by the number of loops that had formed ( n = 3). All values expressed as mean ± SD. All data are representative of at least three independent experiments ( n = 3). Statistical significance was determined by ANOVA; *p < 0.05, **p < 0.01, ***p < 0.001, and ****p < 0.0001.

Article Snippet: To measure in vitro angiogenesis, 48‐well plates were coated with 60 μL of growth factor reduced Matrigel (Corning; 356230) and incubated at 37°C for 30 min. P4 HUVECs were then seeded at 35,000 cells/well with either endothelial growth media (PromoCell; C‐22121) with 1% penicillin–streptomycin (positive control), endothelial basal media (negative control), or endothelial basal media (PromoCell; C‐22221) with 0.1% FBS and 1% penicillin–streptomycin with 5E9 EVs/mL.

Techniques: Isolation, Microscopy

Journal: Bioengineering & Translational Medicine

Article Title: Mesenchymal stem cell extracellular vesicle vascularization bioactivity and production yield are responsive to cell culture substrate stiffness

doi: 10.1002/btm2.10743

Figure Lengend Snippet: Softer 184:527 PDMS substrates improve the angiogenic bioactivity of BM‐MSC EVs. (a) EV production quantified as EV per cell from BM‐MSCs seeded on each substrate made with different ratios of Sylgard 184 and Sylgard 527 ( n = 2). EVs used for this data were from 1 day of collection and isolated and counted separately from the conditioned media from the other 2 days. After media collection, cells were trypsinized and counted. (b) After a scratch was induced, HUVECs were treated with BM‐MSC EVs from the different substrates or growth or basal media, and percent gap closure after 20 h was evaluated via microscopy ( n = 3). (c) HUVECs were resuspended in the different EV treatments or growth or basal endothelial basal media, and tube formation after 3–6 h was quantified by the number of loops that had formed ( n = 3). All values expressed as mean ± SD. Statistical significance was determined by ANOVA; * p < 0.05, ** p < 0.01, and **** p < 0.0001.

Article Snippet: To measure in vitro angiogenesis, 48‐well plates were coated with 60 μL of growth factor reduced Matrigel (Corning; 356230) and incubated at 37°C for 30 min. P4 HUVECs were then seeded at 35,000 cells/well with either endothelial growth media (PromoCell; C‐22121) with 1% penicillin–streptomycin (positive control), endothelial basal media (negative control), or endothelial basal media (PromoCell; C‐22221) with 0.1% FBS and 1% penicillin–streptomycin with 5E9 EVs/mL.

Techniques: Isolation, Microscopy