Limits...
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.

Show MeSH

Related in: MedlinePlus

Tubule formation by HPTCs on 2D surfaces and in 3D gels. (A)–(D) show tubule formation by HPTCs growing on matrigel-coated bottoms of 24-well plate wells. (A) First, a confluent monolayer is formed. (B) Subsequently, the monolayer retracts on one side. (C) Then the monolayer retracts on both sides of a myofibroblast aggregate. (D) Finally, a tubule attached to myofibroblast aggregates is formed. The process is similar to that shown in Figure 1. (E)–(H) show tubule formation by HPTCs suspended in matrigel. (E) Initially, single cells or small groups of cells are present. Note that most of these structures distributed in the 3D gel are out of focus, if a given field is imaged and appear as blurred rings on the images. (F, G) Cell outgrowth occurs (no cyst formation before cell outgrowth), leading to the formation of elongated cords or tubules. The tip cells are typically branched and display multiple filopodia (shown as enlarged in the insets; the branched cell shown in (F) appears blurred due to problems with imaging these structures within the gel). (H) Finally, thin tubules displaying multiple branches are formed. The size of tubules formed in matrigel is typically less than 1 mm, and the tubules are not attached to myofibroblast aggregates (note the different morphology of the structures shown in D and H). Scale bars: (A–D) 1 mm, (E–G) 100 μm and (H) 500 μm.
© Copyright Policy
Related In: Results  -  Collection


getmorefigures.php?uid=PMC4373329&req=5

fig05: Tubule formation by HPTCs on 2D surfaces and in 3D gels. (A)–(D) show tubule formation by HPTCs growing on matrigel-coated bottoms of 24-well plate wells. (A) First, a confluent monolayer is formed. (B) Subsequently, the monolayer retracts on one side. (C) Then the monolayer retracts on both sides of a myofibroblast aggregate. (D) Finally, a tubule attached to myofibroblast aggregates is formed. The process is similar to that shown in Figure 1. (E)–(H) show tubule formation by HPTCs suspended in matrigel. (E) Initially, single cells or small groups of cells are present. Note that most of these structures distributed in the 3D gel are out of focus, if a given field is imaged and appear as blurred rings on the images. (F, G) Cell outgrowth occurs (no cyst formation before cell outgrowth), leading to the formation of elongated cords or tubules. The tip cells are typically branched and display multiple filopodia (shown as enlarged in the insets; the branched cell shown in (F) appears blurred due to problems with imaging these structures within the gel). (H) Finally, thin tubules displaying multiple branches are formed. The size of tubules formed in matrigel is typically less than 1 mm, and the tubules are not attached to myofibroblast aggregates (note the different morphology of the structures shown in D and H). Scale bars: (A–D) 1 mm, (E–G) 100 μm and (H) 500 μm.

Mentions: In order to address tubule formation by HPTCs in 3D gels, we cultured the cells in matrigel. Tubule formation in matrigel (Fig. 5E–H) involved branching of cells at the initial stages of tubule formation and outgrowth of branched cells. Outgrowing branches then formed tubules, and budding from these tubules could occur, giving rise to branched tubular structures (Fig. 5H). It was important to note that formation of epithelial monolayers and coordinated movements of large numbers of cells were not involved in tubule formation in 3D gels. The resulting human proximal tubules obtained in 3D gels were relatively small, displayed multiple branches (Fig. 5H), could be convoluted (Fig. S2), and were never attached to myofibroblast aggregates, in contrast to the tubules obtained on 2D surfaces.


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 by HPTCs on 2D surfaces and in 3D gels. (A)–(D) show tubule formation by HPTCs growing on matrigel-coated bottoms of 24-well plate wells. (A) First, a confluent monolayer is formed. (B) Subsequently, the monolayer retracts on one side. (C) Then the monolayer retracts on both sides of a myofibroblast aggregate. (D) Finally, a tubule attached to myofibroblast aggregates is formed. The process is similar to that shown in Figure 1. (E)–(H) show tubule formation by HPTCs suspended in matrigel. (E) Initially, single cells or small groups of cells are present. Note that most of these structures distributed in the 3D gel are out of focus, if a given field is imaged and appear as blurred rings on the images. (F, G) Cell outgrowth occurs (no cyst formation before cell outgrowth), leading to the formation of elongated cords or tubules. The tip cells are typically branched and display multiple filopodia (shown as enlarged in the insets; the branched cell shown in (F) appears blurred due to problems with imaging these structures within the gel). (H) Finally, thin tubules displaying multiple branches are formed. The size of tubules formed in matrigel is typically less than 1 mm, and the tubules are not attached to myofibroblast aggregates (note the different morphology of the structures shown in D and H). Scale bars: (A–D) 1 mm, (E–G) 100 μm and (H) 500 μm.
© Copyright Policy
Related In: Results  -  Collection

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

fig05: Tubule formation by HPTCs on 2D surfaces and in 3D gels. (A)–(D) show tubule formation by HPTCs growing on matrigel-coated bottoms of 24-well plate wells. (A) First, a confluent monolayer is formed. (B) Subsequently, the monolayer retracts on one side. (C) Then the monolayer retracts on both sides of a myofibroblast aggregate. (D) Finally, a tubule attached to myofibroblast aggregates is formed. The process is similar to that shown in Figure 1. (E)–(H) show tubule formation by HPTCs suspended in matrigel. (E) Initially, single cells or small groups of cells are present. Note that most of these structures distributed in the 3D gel are out of focus, if a given field is imaged and appear as blurred rings on the images. (F, G) Cell outgrowth occurs (no cyst formation before cell outgrowth), leading to the formation of elongated cords or tubules. The tip cells are typically branched and display multiple filopodia (shown as enlarged in the insets; the branched cell shown in (F) appears blurred due to problems with imaging these structures within the gel). (H) Finally, thin tubules displaying multiple branches are formed. The size of tubules formed in matrigel is typically less than 1 mm, and the tubules are not attached to myofibroblast aggregates (note the different morphology of the structures shown in D and H). Scale bars: (A–D) 1 mm, (E–G) 100 μm and (H) 500 μm.
Mentions: In order to address tubule formation by HPTCs in 3D gels, we cultured the cells in matrigel. Tubule formation in matrigel (Fig. 5E–H) involved branching of cells at the initial stages of tubule formation and outgrowth of branched cells. Outgrowing branches then formed tubules, and budding from these tubules could occur, giving rise to branched tubular structures (Fig. 5H). It was important to note that formation of epithelial monolayers and coordinated movements of large numbers of cells were not involved in tubule formation in 3D gels. The resulting human proximal tubules obtained in 3D gels were relatively small, displayed multiple branches (Fig. 5H), could be convoluted (Fig. S2), and were never attached to myofibroblast aggregates, in contrast to the tubules obtained on 2D surfaces.

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