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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

Tubule formation is associated with rapid cell movements. The panels show the same area imaged by differential interference contrast microscopy at consecutive time-points (minutes and seconds are indicated in the lower left corner). The images show living HPTCs in cell culture medium on the bottom of a well of a 24-well plate. The imaged area contains part of a myofibroblast aggregate (right edge). The monolayer has already retracted on one side of the myofibroblast aggregate (note that the upper right area is devoid of cells). Cells are in the process of retracting from the other side and folding up the cell stripe into a tubule. The cell stripe is substantially narrowed over the period of 5 min., as indicated for one region marked by the small arrowheads. A tubule-like structure with two clear borders (large arrowheads) is visible at the end of the observation period, but not at the previous time-points. Thus, this structure and its lower border (marked by the lower large arrowhead) are formed in ∼5 min. Cells at the borders of the stripe (marked by arrow) are quickly integrated into the tubule that is being formed. The dark line on the left side of the panels belongs to a grid, which has been drawn on the outer surface of the well bottom to facilitate spatial orientation during the imaging of cell movements. Scale bar: 200 μm.
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fig02: Tubule formation is associated with rapid cell movements. The panels show the same area imaged by differential interference contrast microscopy at consecutive time-points (minutes and seconds are indicated in the lower left corner). The images show living HPTCs in cell culture medium on the bottom of a well of a 24-well plate. The imaged area contains part of a myofibroblast aggregate (right edge). The monolayer has already retracted on one side of the myofibroblast aggregate (note that the upper right area is devoid of cells). Cells are in the process of retracting from the other side and folding up the cell stripe into a tubule. The cell stripe is substantially narrowed over the period of 5 min., as indicated for one region marked by the small arrowheads. A tubule-like structure with two clear borders (large arrowheads) is visible at the end of the observation period, but not at the previous time-points. Thus, this structure and its lower border (marked by the lower large arrowhead) are formed in ∼5 min. Cells at the borders of the stripe (marked by arrow) are quickly integrated into the tubule that is being formed. The dark line on the left side of the panels belongs to a grid, which has been drawn on the outer surface of the well bottom to facilitate spatial orientation during the imaging of cell movements. Scale bar: 200 μm.

Mentions: Cell movements involved in monolayer reorganization and tubule formation were not only highly coordinated, but also rapid. Figure 2 shows that condensation and folding up of the cell stripe, which ultimately led to tubule formation, occurred within minutes. We were not able to determine definitively how the tubule was folded from the stripe. This would require extensive live cell tracking and detailed analyses of fast movements of large numbers of cells. Such extensive live cell analyses would be performed in a separate study.


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)

Tubule formation is associated with rapid cell movements. The panels show the same area imaged by differential interference contrast microscopy at consecutive time-points (minutes and seconds are indicated in the lower left corner). The images show living HPTCs in cell culture medium on the bottom of a well of a 24-well plate. The imaged area contains part of a myofibroblast aggregate (right edge). The monolayer has already retracted on one side of the myofibroblast aggregate (note that the upper right area is devoid of cells). Cells are in the process of retracting from the other side and folding up the cell stripe into a tubule. The cell stripe is substantially narrowed over the period of 5 min., as indicated for one region marked by the small arrowheads. A tubule-like structure with two clear borders (large arrowheads) is visible at the end of the observation period, but not at the previous time-points. Thus, this structure and its lower border (marked by the lower large arrowhead) are formed in ∼5 min. Cells at the borders of the stripe (marked by arrow) are quickly integrated into the tubule that is being formed. The dark line on the left side of the panels belongs to a grid, which has been drawn on the outer surface of the well bottom to facilitate spatial orientation during the imaging of cell movements. Scale bar: 200 μm.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC4373329&req=5

fig02: Tubule formation is associated with rapid cell movements. The panels show the same area imaged by differential interference contrast microscopy at consecutive time-points (minutes and seconds are indicated in the lower left corner). The images show living HPTCs in cell culture medium on the bottom of a well of a 24-well plate. The imaged area contains part of a myofibroblast aggregate (right edge). The monolayer has already retracted on one side of the myofibroblast aggregate (note that the upper right area is devoid of cells). Cells are in the process of retracting from the other side and folding up the cell stripe into a tubule. The cell stripe is substantially narrowed over the period of 5 min., as indicated for one region marked by the small arrowheads. A tubule-like structure with two clear borders (large arrowheads) is visible at the end of the observation period, but not at the previous time-points. Thus, this structure and its lower border (marked by the lower large arrowhead) are formed in ∼5 min. Cells at the borders of the stripe (marked by arrow) are quickly integrated into the tubule that is being formed. The dark line on the left side of the panels belongs to a grid, which has been drawn on the outer surface of the well bottom to facilitate spatial orientation during the imaging of cell movements. Scale bar: 200 μm.
Mentions: Cell movements involved in monolayer reorganization and tubule formation were not only highly coordinated, but also rapid. Figure 2 shows that condensation and folding up of the cell stripe, which ultimately led to tubule formation, occurred within minutes. We were not able to determine definitively how the tubule was folded from the stripe. This would require extensive live cell tracking and detailed analyses of fast movements of large numbers of cells. Such extensive live cell analyses would be performed in a separate study.

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