Tisseel kit

Manufactured by Baxter

Sourced in United Kingdom, Australia

The Tisseel Kit is a surgical sealant product that contains a sealing protein and a thrombin solution. It is designed to be used as an adjunct to standard surgical techniques for the sealing of bleeding and fluid leaks.

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Lab products found in correlation

Dulbecco modified eagle medium (dmem), by Corning (1 mentions) Phosphate buffered saline (pbs), by Thermo Fisher Scientific (1 mentions) Trasylol, by Bayer (1 mentions) Fibroblast growth factor 2 (fgf2), by Miltenyi Biotec (1 mentions) Vascular endothelial growth factor (vegf), by Miltenyi Biotec (1 mentions) Sdf 1α, by Miltenyi Biotec (1 mentions) Egm 2, by PromoCell (1 mentions)

Close spelling variants of this product

Tisseel (71 citations) Tisseel Kit VH 1.0 (2 citations)

13 protocols using tisseel kit

Tissue Repair Exosome Release Assay

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For the Tisseel release assay, a Tisseel kit (Baxter, #1504517) was prepared by adding 1 mL of Fibrinolysis Inhibitor to Sealer Protein Concentrate and adding 1 mL of CaCl₂ Solution to Thrombin 500. Both solutions were incubated for 20 min at 37 °C. One vial of PEP (Lot# 19004-B1) was resuspended with 400 uL heparin and 1.25 mL deionized water. The entire PEP solution was added to the Fibrinolysis Inhibitor/Sealer Protein Concentrate Solution. This solution was combined in each well of a 12-well plate in a 2:1 ratio with the CaCl₂/Thrombin 500 solution for a total volume of 990 ul per well.
For the Collagen release assay, one vial of PEP (Lot# 19004-B1) was reconstituted in 2.5 mL of sterile water and filtered through a 0.22 um filter. Five hundred microliters PEP was combined with 500 uL of 6 mg/mL collagen (Collagen Solutions FS22004). NaOH was added to a final concentration of 0.02 M. The entire 1 mL solution was added to a single well of a 12-well plate and incubated at 37 °C until solid. The final concentration of PEP for both assays in each well was 1 × 10¹² exosomes/mL. One mL of serum-free Dulbecco’s Modified Eagle Medium 1× (DMEM, Corning, 10-013CV) was added to each well. All media was collected daily and replaced with another 1 mL of serum-free DMEM for the time-course of the experiment.

Rolland T.J., Peterson T.E., Singh R.D., Rizzo S.A., Boroumand S., Shi A., Witt T.A., Nagel M., Kisby C.K., Park S., Rowe L.A., Paradise C.R., Becher L.R., Paradise B.D., Stalboerger P.G., Trabuco E.C, & Behfar A. (2022). Exosome biopotentiated hydrogel restores damaged skeletal muscle in a porcine model of stress urinary incontinence. NPJ Regenerative Medicine, 7, 58.

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Stabilized PEP-Loaded Fibrin Sealant for Tissue Regeneration

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PEP was obtained from the API at the Mayo Clinic Center for Regenerative Medicine. The product was formulated and stored in a stabilized lyophilized powder form in vials to allow for room temperature storage until processing (Fig. 2A).
Fibrin sealant is a biodegradable pulp-like tissue that can be used as a drug delivery vehicle, and is very effective at achieving a local and sustained release of exosomes. Before preparation of the TISSEEL kit (Baxter International), a vial of sealed PEP powder was mixed with 1 mL PBS (Gibco) to prepare the 100 % (vol/vol) PEP solution. We added 400 μL PEP solution into the 600 μL CaCl₂ solution (one of the contents of the TISSEEL kit), and the solution was normalized to a 40 % (vol/vol) concentration. Manufacturer directions were then followed to finish kit preparation (Fig. 2B). The final concentration of PEP in TISSEEL was 20 % (vol/vol). The gel was manually cut into small cubes (3 × 3 × 3 mm). In the co-culture model, a single cube was placed into the small hole in one well of a 6-well plate. The culture medium with PEP gel was used to simulate the PEP microenvironment in vivo. For osteogenic induction, the medium with PEP as described above was used as the positive control group. For the in vivo trial, the cube was placed directly on the RC repair site, between the supraspinatus tendon and the greater tuberosity.

Ren Y., Zhang S., Wang Y., Jacobson D.S., Reisdorf R.L., Kuroiwa T., Behfar A., Moran S.L., Steinmann S.P, & Zhao C. (2021). Effects of purified exosome product on rotator cuff tendon-bone healing in vitro and in vivo. Biomaterials, 276, 121019.

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In Vivo Osteogenic Potential of Nucleofected MSCs

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To evaluate the in vivo osteogenic potential of the cells after nucleofection, we first anesthetized immunodeficient (NOD/SCID) mice by administering 2%–3% isoflurane and placing the animals on a 37°C heating pad. Aliquots of 3 × 10⁶ cells were resuspended in 50 µl fibrin gel (Tisseel kit; Baxter, Vienna, Austria, http://www.baxter.com) and injected into the thigh muscles of the animals (n = 5). Each mouse received one injection of MSCs (aiMSCs, tiMSCs, or BMSCs) that had been nucleofected with BMP6. Bone formation was evaluated after 4 weeks using micro-computed tomography (µCT). The mice were euthanized, and their limbs were harvested for histologic analysis.

Sheyn D., Ben-David S., Shapiro G., De Mel S., Bez M., Ornelas L., Sahabian A., Sareen D., Da X., Pelled G., Tawackoli W., Liu Z., Gazit D, & Gazit Z. (2016). Human Induced Pluripotent Stem Cells Differentiate Into Functional Mesenchymal Stem Cells and Repair Bone Defects. Stem Cells Translational Medicine, 5(11), 1447-1460.

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Fibrin-Based Engineered Skin Grafts

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Transparent fibrin mats were prepared in laminar hood using 2 or 5 mL TISSEEL kit (Baxter). Fibrinogen from the kit was diluted two times above the recommended reconstitution using 1.1% sodium chloride solution (NaCl) containing 1 mM of calcium chloride (CaCl₂); this solution was subsequently mixed in equal volume with thrombin provided by the kit, diluted to 3 IU mL⁻¹ using 1.1% NaCl and 1 mM CaCl₂ solution. The above mixed solutions were dispensed uniformly in 10 × 10 cm² dishes, left at room temperature for 10–15 min for complete polymerization, and stored at 4 °C until use. To prevent fibrinolysis during the culture of HEKs on the fibrin mat, aprotinin (Trasylol, Bayer) was added to a final concentration of 150 kIU mL⁻¹ in the culture medium at each feeding. For transplantation, fibrin mats were either coated with 2.5 μg cm⁻² LN-511 or LN-421 and incubated at 4 °C overnight, or seeded with lethally γ-irradiated 3T3-J2 fibroblasts and incubated at 37 °C overnight. HEKs between passages 1 and 3 were then seeded the next day at 10 000 cells per cm² and grown to confluence. On the day of surgery, all grafts were washed twice with respective serum-free medium (KGM-CD for laminin samples and fresh DMEM for 3T3 co-culture sample).

Tjin M.S., Chua A.W., Moreno-Moral A., Chong L.Y., Tang P.Y., Harmston N.P., Cai Z., Petretto E., Tan B.K, & Tryggvason K. (2018). Biologically relevant laminin as chemically defined and fully human platform for human epidermal keratinocyte culture. Nature Communications, 9, 4432.

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Decellularized Allograft Tissue Engineering

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Structural allografts were harvested from fresh 67 C57BL/6J mouse cadavers using a trephine with inner diameter of 5 mm. Tissue harvest was performed in accordance to the approved IACUC protocol #007961. The allografts were scraped to remove any soft tissue and washed with PBS. For complete decellularization, the allografts were washed for 1 hour in PBS 0.1 wt%/vol% EDTA at room temperature (RT) with rotation, then incubated in the following solutions according to published protocols⁴⁷, ⁴⁸: first: in PBS 0.1 wt%/vol% EDTA and 10 mM Tris overnight at 4°C with rotation; second: PBS 10 mM Tris and 0.5 wt%/vol% sodium dodecyl sulfate for 24 hours at RT; third: enzymatic treatment with PBS 10 mM Tris, 50 U/mL DNase, 1 U/mL RNase for 5 hours at RT with rotation and sterilized with 70% EtOH overnight. Prior to changing the solutions, the allografts were washed for 1 hour with PBS at RT. Grafts were frozen at −80°C prior to implantation and were thawed 24 hours before the surgical procedure by placing them in serum free medium overnight. For cell coating, 2 × 10⁵ BM‐MSC‐Luc2 or iNCC‐MPC‐Luc2 were resuspended in 12.5 μL fibrin gel (Tisseel kit, Baxter). The fibrin‐cell mixture was rapidly added onto the top of the allograft, which was incubated for at least 30 minutes at 37°C prior to implantation to allow gelation and attachment to the graft.

Glaeser J.D., Behrens P., Stefanovic T., Salehi K., Papalamprou A., Tawackoli W., Metzger M.F., Eberlein S., Nelson T., Arabi Y., Kim K., Baloh R.H., Ben‐David S., Cohn‐Schwartz D., Ryu R., Bae H.W., Gazit Z, & Sheyn D. (2021). Neural crest‐derived mesenchymal progenitor cells enhance cranial allograft integration. Stem Cells Translational Medicine, 10(5), 797-809.

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Unilateral Dorsal Rootlet Transection

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Unilateral transection of four dorsal rootlets was carried out as described in our previous study^{34 (link)}. Briefly, a skin incision was made along the dorsal midline under isoflurane anesthesia and analgesia (Vetergesic 0.1-0.05mg/kg; Ceva, France). The prominent T2 process was located and hemilaminectomies were performed from C4 to T2, and the dura was incised with a pair of micro scissors to reveal the dorsal roots. The rootlets of C6, C7, C8, and T1 were transected with micro scissors as close as possible to the spinal cord in a plane approximately perpendicular to their entry into the spinal cord. The cut rootlets were re-apposed and held in place with fibrin glue (Tisseel Kit; Baxter, Thetford, UK).

Minkelyte K., Li D., Li Y, & Ibrahim A. (2023). Transplantation of Cryopreserved Olfactory Ensheathing Cells Restores Loss of Functions in an Experimental Model. Cell Transplantation, 32, 09636897231199319.

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Spinal Cord Injury Repair with bOECs

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The same surgical procedure was carried out as in the control groups. The rats received bOECs encapsulated within a collagen gel immediately after four unilateral dorsal rootlets were transected. The transplant was applied between the cut ends of the rootlets and their original entry point on the spinal cord and held in place with fibrin glue (Tisseel Kit, Baxter, Thetford, UK).

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Transplanting OECs to Repair Dorsal Root Injuries

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The matrix containing cultured OECs expressing GFP was scraped from a Petri dish,
applied between the cut ends of dorsal roots and their original entry points into the
spinal cord, and retained in place with fibrin glue (Tisseel Kit, Baxter, Thetford, UK).
Six rats received the injury alone, i.e., without the OEC transplant and were used as
controls.

Collins A., Ibrahim A., Li D., Liadi M, & Li Y. (2019). Reconstruction of the Damaged Dorsal Root Entry Zone by Transplantation of Olfactory Ensheathing Cells. Cell Transplantation, 28(9-10), 1212-1219.

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Prevascularized Tubular Bone Graft Construct

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Tubular mPCL scaffolds with an internal diameter of 5 mm were fabricated via melt electrowriting and CaP-coated as described above^{5 (link),7 ,25 (link)}. The mPCL-CaP tubular scaffolds were seeded with 100,000 hOB/scaffold in 30 µl serum-free MEMα and cultured in well-plates for 2 weeks in expansion media followed by 7 weeks in osteogenic media. Throughout the course of the scaffold culture, hOBs migrated from the scaffold onto the tissue-culture plastic of the well plates and formed a hOB monolayer. The hOB monolayers were cultured together with the scaffold and were allowed to form a hOB cell sheet.
A prevascularized niche was generated using 4% GelMA-based hydrogels containing HUVECs and MSCs (10:1 ratio; 5.5 × 10⁶ cells/ml total) 7 days before implantation and pre-cultured in EGM2 (PromoCell, Heidelberg, Germany) supplemented with 125 ng/ml SDF-1α, VEGF and FGF2 (Miltenyi Biotec, NSW, Australia). At the time of implantation, the vascular gel was placed inside the tubular hOB scaffold. The hOB cell sheet was mixed with 30 µl fibrin glue (TISSEEL™ kit, Baxter Healthcare, Australia) and 20 µl of rhBMP-2 (1.5 µg/µl; INFUSE^®, Medtronic^{25 (link)}. Two tubular hTEBC were implanted into left and right subcutaneous pockets on the back of male NSG mice as described above.

McGovern J.A., Bock N., Shafiee A., Martine L.C., Wagner F., Baldwin J.G., Landgraf M., Lahr C.A., Meinert C., Williams E.D., Pollock P.M., Denham J., Russell P.J., Risbridger G.P., Clements J.A., Loessner D., Holzapfel B.M, & Hutmacher D.W. (2021). A humanized orthotopic tumor microenvironment alters the bone metastatic tropism of prostate cancer cells. Communications Biology, 4, 1014.

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Subcutaneous Implantation of hTEBC Constructs

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Immediately prior to subcutaneous implantation, the mPCL-CaP in vitro constructs were combined using 60 µl fibrin glue (TISSEEL™ kit, Baxter Healthcare, Australia) embedded with 30 µg rhBMP-7 (Olympus Biotech Corporation, USA). Additionally, the rhBMP-7 and fibrin glue were combined with a hOB cell sheet isolated from the well plates in which the scaffolds were cultured for the hOB laden in vitro constructs. The hOB cell sheet was included as an additional source of hOBs in the hTEBC preparation. The sandwich scaffolds pre-seeded with hOBs were implanted into the left-back of the mouse whereas the unseeded scaffolds were implanted into the right-back of the mouse. To implant the scaffolds, two longitudinal incisions were made on the skin on the left and right back of the mouse. Subcutaneous pockets were created using blunt scissors to gently separate the subcutaneous space. The prepared hTEBCs were inserted into the prepared pockets and the incisions closed with wound closure autoclips (Kent Scientific Corporation, CT, USA). Autoclips were removed by 7−10 days when the surgical site had healed^{5 (link)}.

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