5. Filter.

(a)

0.2 μm injection filter (e.g., B. Braun, Intrapur^®Neonat).

(b)

0.2 μm sterile filter for gas exchange and pressure com-

pensation, connectable to Luer lock fittings.

6. Small diameter vascular grafts to be studied, inner diameter

5.0 mm. Here, we used bacterial nanocellulose small diameter

vascular grafts with and without fibronectin coating.

2.2

Assays and

Staining Solutions

1. WST-1 reagent (Roche, Mannheim, Germany, cat. nr.: 05 015

944 001).

2. Alexa Fluor 488 AcLDL (Thermofisher, Carlsbad, USA, cat.

nr.: L23380).

3. Rhodamine Phalloidin Reagent (Abcam, Cambridge, USA,

cat. nr.: ab235138).

4. NucBlue™Live Ready Probes™Reagent (Invitrogen, Carls-

bad, USA, cat. nr.: R37605).

5. Acridine orange (Invitrogen, Carlsbad, USA, cat. nr.: A1301).

2.3

Cell Culture

1. Endothelial cell line (here, we used rabbit endothelial progeni-

tor cells (rbEPC), isolated from Chinchilla bastard rabbit blood

or human saphenous vein endothelial cells (HSVEC)).

2. Cell line specific cell culture medium containing 10% fetal calf

serum.

3. Consumables for cell culture (flasks, conical tubes, 6-well

plates, serological pipettes, detachment reagent).

2.4

Antibodies

1. CD31/PECAM-1 antibody that fits to your cell line (here, we

used CD31/PECAM-1 mouse anti-human antibody [R&D

Systems, Minneapolis, USA, cat. nr.: BBA7]).

2. Secondary antibody that fits to your primary antibody (here,

we used Donkey anti-mouse Alexa Fluro 488 [Jackson Immu-

noResearch, West Grove, USA, cat. nr.: 715-545-152]).

2.5

Chemicals and

Solutions

1. Phosphate buffered saline without calcium and magnesium

(PBS).

2. PBS with calcium and magnesium (PBS+).

3. PBS with 1% penicillin-streptomycin cell culture supplement.

4. Penicillin-streptomycin cell culture supplement.

5. Normal donkey serum.

6. Paraformaldehyde (PFA) solution, 4%.

7. Bovine serum albumin (BSA).

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Max Wacker et al.