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Generation of easily accessible human kidney tubules on two-dimensional surfaces in vitro.

Zhang H, Lau SF, Heng BF, Teo PY, Alahakoon PK, Ni M, Tasnim F, Ying JY, Zink D - J. Cell. Mol. Med. (2010)

Bottom Line: However, after triggering the process, the formation of renal tubules occurs with remarkable independence from the substrate architecture.Human proximal tubules generated on 2D surfaces typically have a length of several millimetres, and are easily accessible for manipulations and imaging, which makes them attractive for basic research and in vitro nephrotoxicology.The experimental system described also allows for in vitro studies on how primary human kidney cells regenerate renal structures after organ disruption.

View Article: PubMed Central - PubMed

Affiliation: Institute of Bioengineering and Nanotechnology, The Nanos, Singapore, Singapore.

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Related in: MedlinePlus

Triggering of tubulogenesis in the presence of a 3D substrate architecture. HPTCs were grown to confluency on glass cover slips (A, D and G), in the wells of 24-well plates consisting of tissue culture plastic (B, E and H), and in the wells of diagnostic printed slides (C, F and I). Cover slips with a side length of 18 mm are used. The wells of 24-well plates and diagnostic printed slides are 15 mm and 2 mm in diameter, respectively. Cells on the different devices are monitored over a time period of 8 days. (A)–(C) show the confluent monolayers at day 2. The edges of the different substrates used are indicated by large arrowheads. (E, F) Monolayer retraction starts at day 3 at the edges of the wells (marked by large arrowheads) of 24-well plates and diagnostic printed slides. This leads to areas devoid of cells (marked by a small arrowhead in F). No rearrangements are observed at (D) day 3 and (G) day 8 at the edges of cover slips (marked by large arrowheads), which do not have a 3D structure. The monolayer is still intact on cover slips. By contrast, major rearrangements are noted at day 8 in the wells of (H) 24-well plates and (I) diagnostic printed slides. Formation of tubules (marked by small arrowhead in H) and myofibroblast aggregates (marked by small arrowhead in I) is observed. The wells of 24-well plates and diagnostic printed slides provide different surface chemistries and surface areas. However, in both cases, the edge is a 3D structure, in contrast to the edge of cover slips. Scale bar: 500 μm.
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fig07: Triggering of tubulogenesis in the presence of a 3D substrate architecture. HPTCs were grown to confluency on glass cover slips (A, D and G), in the wells of 24-well plates consisting of tissue culture plastic (B, E and H), and in the wells of diagnostic printed slides (C, F and I). Cover slips with a side length of 18 mm are used. The wells of 24-well plates and diagnostic printed slides are 15 mm and 2 mm in diameter, respectively. Cells on the different devices are monitored over a time period of 8 days. (A)–(C) show the confluent monolayers at day 2. The edges of the different substrates used are indicated by large arrowheads. (E, F) Monolayer retraction starts at day 3 at the edges of the wells (marked by large arrowheads) of 24-well plates and diagnostic printed slides. This leads to areas devoid of cells (marked by a small arrowhead in F). No rearrangements are observed at (D) day 3 and (G) day 8 at the edges of cover slips (marked by large arrowheads), which do not have a 3D structure. The monolayer is still intact on cover slips. By contrast, major rearrangements are noted at day 8 in the wells of (H) 24-well plates and (I) diagnostic printed slides. Formation of tubules (marked by small arrowhead in H) and myofibroblast aggregates (marked by small arrowhead in I) is observed. The wells of 24-well plates and diagnostic printed slides provide different surface chemistries and surface areas. However, in both cases, the edge is a 3D structure, in contrast to the edge of cover slips. Scale bar: 500 μm.

Mentions: In order to determine whether the presence of a 3D edge indeed enhanced the initiation of tubule formation, we seeded cells in parallel on 18 mm cover slips (no 3D edge), 24-well cell culture plates (with 3D edge) and diagnostic printed slides (with 3D edge). Figure 7 shows that initial retraction of the epithelium first occurred at the edges of the wells of the 24-well plates (well diameter = 15 mm) and diagnostic printed slides (well diameter = 2 mm). Subsequently, monolayer reorganization as well as cell aggregate and tubule formation were observed within the wells of these devices. By contrast, the epithelial monolayer was not reorganized on cover slips during the monitoring period of 8 days, and no retraction of the monolayer occurred at the edges of the cover slips (Fig. 7).


Generation of easily accessible human kidney tubules on two-dimensional surfaces in vitro.

Zhang H, Lau SF, Heng BF, Teo PY, Alahakoon PK, Ni M, Tasnim F, Ying JY, Zink D - J. Cell. Mol. Med. (2010)

Triggering of tubulogenesis in the presence of a 3D substrate architecture. HPTCs were grown to confluency on glass cover slips (A, D and G), in the wells of 24-well plates consisting of tissue culture plastic (B, E and H), and in the wells of diagnostic printed slides (C, F and I). Cover slips with a side length of 18 mm are used. The wells of 24-well plates and diagnostic printed slides are 15 mm and 2 mm in diameter, respectively. Cells on the different devices are monitored over a time period of 8 days. (A)–(C) show the confluent monolayers at day 2. The edges of the different substrates used are indicated by large arrowheads. (E, F) Monolayer retraction starts at day 3 at the edges of the wells (marked by large arrowheads) of 24-well plates and diagnostic printed slides. This leads to areas devoid of cells (marked by a small arrowhead in F). No rearrangements are observed at (D) day 3 and (G) day 8 at the edges of cover slips (marked by large arrowheads), which do not have a 3D structure. The monolayer is still intact on cover slips. By contrast, major rearrangements are noted at day 8 in the wells of (H) 24-well plates and (I) diagnostic printed slides. Formation of tubules (marked by small arrowhead in H) and myofibroblast aggregates (marked by small arrowhead in I) is observed. The wells of 24-well plates and diagnostic printed slides provide different surface chemistries and surface areas. However, in both cases, the edge is a 3D structure, in contrast to the edge of cover slips. Scale bar: 500 μm.
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Related In: Results  -  Collection

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fig07: Triggering of tubulogenesis in the presence of a 3D substrate architecture. HPTCs were grown to confluency on glass cover slips (A, D and G), in the wells of 24-well plates consisting of tissue culture plastic (B, E and H), and in the wells of diagnostic printed slides (C, F and I). Cover slips with a side length of 18 mm are used. The wells of 24-well plates and diagnostic printed slides are 15 mm and 2 mm in diameter, respectively. Cells on the different devices are monitored over a time period of 8 days. (A)–(C) show the confluent monolayers at day 2. The edges of the different substrates used are indicated by large arrowheads. (E, F) Monolayer retraction starts at day 3 at the edges of the wells (marked by large arrowheads) of 24-well plates and diagnostic printed slides. This leads to areas devoid of cells (marked by a small arrowhead in F). No rearrangements are observed at (D) day 3 and (G) day 8 at the edges of cover slips (marked by large arrowheads), which do not have a 3D structure. The monolayer is still intact on cover slips. By contrast, major rearrangements are noted at day 8 in the wells of (H) 24-well plates and (I) diagnostic printed slides. Formation of tubules (marked by small arrowhead in H) and myofibroblast aggregates (marked by small arrowhead in I) is observed. The wells of 24-well plates and diagnostic printed slides provide different surface chemistries and surface areas. However, in both cases, the edge is a 3D structure, in contrast to the edge of cover slips. Scale bar: 500 μm.
Mentions: In order to determine whether the presence of a 3D edge indeed enhanced the initiation of tubule formation, we seeded cells in parallel on 18 mm cover slips (no 3D edge), 24-well cell culture plates (with 3D edge) and diagnostic printed slides (with 3D edge). Figure 7 shows that initial retraction of the epithelium first occurred at the edges of the wells of the 24-well plates (well diameter = 15 mm) and diagnostic printed slides (well diameter = 2 mm). Subsequently, monolayer reorganization as well as cell aggregate and tubule formation were observed within the wells of these devices. By contrast, the epithelial monolayer was not reorganized on cover slips during the monitoring period of 8 days, and no retraction of the monolayer occurred at the edges of the cover slips (Fig. 7).

Bottom Line: However, after triggering the process, the formation of renal tubules occurs with remarkable independence from the substrate architecture.Human proximal tubules generated on 2D surfaces typically have a length of several millimetres, and are easily accessible for manipulations and imaging, which makes them attractive for basic research and in vitro nephrotoxicology.The experimental system described also allows for in vitro studies on how primary human kidney cells regenerate renal structures after organ disruption.

View Article: PubMed Central - PubMed

Affiliation: Institute of Bioengineering and Nanotechnology, The Nanos, Singapore, Singapore.

Show MeSH
Related in: MedlinePlus