bovine telocol 6 type i collagen solution  (Advanced Biomatrix Inc)

Journal: STAR Protocols

Article Title: Protocol for generating and analyzing organ-on-chip using human and mouse intestinal organoids

doi: 10.1016/j.xpro.2024.103037

Figure Lengend Snippet: Graphical illustration of chip layout and preparation One chip consists of 9 wells of the OrganoPlate 3-lane 40. Chip layout shows the top channel with the top inlet (A1) and top outlet (A3), the gel channel with the gel inlet (B1) and gel outlet (B3) and the bottom channel with the bottom inlet (C1) and bottom outlet (C3). All in- and outlets are marked by a green rectangle. The chip itself is located in B2 and marked by the pink square. The chip preparation is prepared by the addition of Collagen I to the gel channel (i), then Collagen IV is added to the top channel as a coating (ii) before medium and cell suspension are added in the last step (iii). Collagen I in green, air as light blue, spacer in gray, coating solution in orange and medium in pink.

Article Snippet: Bovine collagen I (TeloCol-6 bovine type I collagen solution, 6 mg/mL) , Advanced Biomatrix , Cat#5225.

Techniques: Suspension

Journal: STAR Protocols

Article Title: Protocol for generating and analyzing organ-on-chip using human and mouse intestinal organoids

doi: 10.1016/j.xpro.2024.103037

Figure Lengend Snippet: Image of chip appearance during the preparation process (A) Images show the appearance of the chip during the preparation process. Left image: Empty chip before starting. Middle image: Chip after ECM addition marked by two dark lines (pink arrow heads). Right image: Chip with ECM after addition of coating solution marked by the disappearance of one dark line. (B) Brightfield image of chip during coating step. Middle channel contains Collagen I that presents with structured texture (∗). The top channel contains coating solution. Bottom channel is empty, thus showing the meniscus of the Collagen in the middle channel (arrow). Scale bar 500 μm.

Article Snippet: Bovine collagen I (TeloCol-6 bovine type I collagen solution, 6 mg/mL) , Advanced Biomatrix , Cat#5225.

Techniques:

Journal: STAR Protocols

Article Title: Protocol for generating and analyzing organ-on-chip using human and mouse intestinal organoids

doi: 10.1016/j.xpro.2024.103037

Figure Lengend Snippet: Graphical illustration of single cell seeding Organoids are dissociated, and single cells (dark red) seeded into the top channel of the chip. Cells are allowed to settle against Collagen I (green) to facilitate cell attachment. Details are given in the text. Medium is shown in pink.

Article Snippet: Bovine collagen I (TeloCol-6 bovine type I collagen solution, 6 mg/mL) , Advanced Biomatrix , Cat#5225.

Techniques: Cell Attachment Assay

Journal: STAR Protocols

Article Title: Protocol for generating and analyzing organ-on-chip using human and mouse intestinal organoids

doi: 10.1016/j.xpro.2024.103037

Figure Lengend Snippet: Displacement of ECM during preparation step Collagen I was displaced from the middle to the top channel (white arrows) due to incomplete polymerization.

Article Snippet: Bovine collagen I (TeloCol-6 bovine type I collagen solution, 6 mg/mL) , Advanced Biomatrix , Cat#5225.

Techniques:

Journal: STAR Protocols

Article Title: Protocol for generating and analyzing organ-on-chip using human and mouse intestinal organoids

doi: 10.1016/j.xpro.2024.103037

Figure Lengend Snippet: Graphical illustration of barrier formation and chip cross section The epithelial barrier (dark red) forms against the Collagen I (green) over time with constant flow and regular medium (pink) changes. The cross section shows the side view (YZ) and the focus on the barrier at the cell-collagen interface (right). It is a commonly used z plane to assess the barrier integrity.

Article Snippet: Bovine collagen I (TeloCol-6 bovine type I collagen solution, 6 mg/mL) , Advanced Biomatrix , Cat#5225.

Techniques:

Journal: STAR Protocols

Article Title: Protocol for generating and analyzing organ-on-chip using human and mouse intestinal organoids

doi: 10.1016/j.xpro.2024.103037

Figure Lengend Snippet: Human organoid growth on the chip tract over time (A) Brightfield images showing the barrier formation at the level of the Collagen I on day 3, 6, 8 and 10. (B) Brightfield images showing the barrier formation at the top of the chip on day 3, 6, 8 and 10. Scale bar 500 μm and 200 μm (close-up). d = days post seeding.

Article Snippet: Bovine collagen I (TeloCol-6 bovine type I collagen solution, 6 mg/mL) , Advanced Biomatrix , Cat#5225.

Techniques:

Journal: STAR Protocols

Article Title: Protocol for generating and analyzing organ-on-chip using human and mouse intestinal organoids

doi: 10.1016/j.xpro.2024.103037

Figure Lengend Snippet: Permeability assay of mouse OoC (A) Bright-field image and graphical illustration of an OoC of mouse distal colon with a leaky barrier. The epithelial cells (dark red) form a barrier against the Collagen I (light green) which inhibits the translocation of dye (green) from the top channel to the bottom channel containing medium (pink). (B) Permeability assay using FITC-dextran 4 kDa (green) and TRITC-dextran 70 kDa (yellow) assessing leakiness initially (t0) and after 30 min (t1) of OoC depicted in A. Arrow indicates leakiness. (C) Brightfield image and graphical illustration of an OoC of mouse distal colon with an impermeable barrier. (D) Permeability assay using FITC-dextran 4 kDa (green) and TRITC-dextran 70 kDa (yellow) assessing leakiness initially (t0) and after 30 min (t1) of OoC depicted in C. Scale bar 500 μm.

Article Snippet: Bovine collagen I (TeloCol-6 bovine type I collagen solution, 6 mg/mL) , Advanced Biomatrix , Cat#5225.

Techniques: Permeability, Translocation Assay

Journal: STAR Protocols

Article Title: Protocol for generating and analyzing organ-on-chip using human and mouse intestinal organoids

doi: 10.1016/j.xpro.2024.103037

Figure Lengend Snippet: Staining of human OoC (A) 3D reconstruction of human colon OoC showing intact barrier at the cell-Collagen I interface. (B) Images show the epithelial barrier formed at the cell-Collagen I interface (arrow) from different angles. The main image shows the XY orientation. The lower panel shows the XZ and the right panel the YZ view. The yellow crosshairs show the location of the optical cut. Scale bar 500 μm. (C) Graphical representation of the crypt-villus structure present in vivo . Brightfield and confocal microscopy image showing the crypt-villus structure formed at the cell-Collagen I interface in the OoC. Scale bar 200 μm and 100 μm respectively. Optical cross-section showing of invaginations of the cell monolayer and the presence of ZO-1 marking the apical surface. The main image shows the XY orientation. The lower panel shows the XZ and the right panel the YZ view, the yellow crosshairs the location of the optical cut. Scale bar 20 μm. (D) Orthogonal cross section of a z-stack showing the polarization of the cells with stress fibers (left image) as sign for cell attachment, nuclei within the cells (middle image) and microvilli structures towards the apical side (right image). XZ and YZ projections depict the basal and apical (∗) polarization of the cells with shedding cells on the apical side marked by arrow heads. The plot shows the mean intensity (actin) of each z-stack. Scale bar 25 μm. (E) The basal side (b) containing stress fibers shows a trend towards lower actin intensity while the apical side (a) shows an enrichment of actin which is caused by dense microvilli coverage. In all images actin in shown in magenta and nuclei in cyan. The right image in panel C shows tight junction protein ZO-1 in gray.

Article Snippet: Bovine collagen I (TeloCol-6 bovine type I collagen solution, 6 mg/mL) , Advanced Biomatrix , Cat#5225.

Techniques: Staining, In Vivo, Confocal Microscopy, Cell Attachment Assay

Journal: STAR Protocols

Article Title: Protocol for generating and analyzing organ-on-chip using human and mouse intestinal organoids

doi: 10.1016/j.xpro.2024.103037

Figure Lengend Snippet:

Article Snippet: Bovine collagen I (TeloCol-6 bovine type I collagen solution, 6 mg/mL) , Advanced Biomatrix , Cat#5225.

Techniques: Recombinant, Derivative Assay, Stable Transfection, Transfection, Software, Adhesive, Transferring, Microscopy, Fluorescence, Sterility

Journal: STAR Protocols

Article Title: Protocol for generating and analyzing organ-on-chip using human and mouse intestinal organoids

doi: 10.1016/j.xpro.2024.103037

Figure Lengend Snippet: Human ECM mix

Article Snippet: Bovine collagen I (TeloCol-6 bovine type I collagen solution, 6 mg/mL) , Advanced Biomatrix , Cat#5225.

Techniques: Concentration Assay, Neutralization

Journal: STAR Protocols

Article Title: Protocol for generating and analyzing organ-on-chip using human and mouse intestinal organoids

doi: 10.1016/j.xpro.2024.103037

Figure Lengend Snippet: Murine ECM mix

Article Snippet: Bovine collagen I (TeloCol-6 bovine type I collagen solution, 6 mg/mL) , Advanced Biomatrix , Cat#5225.

Techniques: Concentration Assay