Limits...
Generation of an expandable intermediate mesoderm restricted progenitor cell line from human pluripotent stem cells.

Kumar N, Richter J, Cutts J, Bush KT, Trujillo C, Nigam SK, Gaasterland T, Brafman D, Willert K - Elife (2015)

Bottom Line: One way to mitigate this risk is to develop expandable progenitor cell populations with restricted differentiation potential.Here, we used a cellular microarray technology to identify a defined and optimized culture condition that supports the derivation and propagation of a cell population with mesodermal properties.Interestingly, IMP cells fail to differentiate into other mesodermally-derived tissues, including blood and heart, suggesting that these cells are restricted to an intermediate mesodermal fate.

View Article: PubMed Central - PubMed

Affiliation: Department of Cellular and Molecular Medicine, University of California, San Diego, San Diego, United States.

ABSTRACT
The field of tissue engineering entered a new era with the development of human pluripotent stem cells (hPSCs), which are capable of unlimited expansion whilst retaining the potential to differentiate into all mature cell populations. However, these cells harbor significant risks, including tumor formation upon transplantation. One way to mitigate this risk is to develop expandable progenitor cell populations with restricted differentiation potential. Here, we used a cellular microarray technology to identify a defined and optimized culture condition that supports the derivation and propagation of a cell population with mesodermal properties. This cell population, referred to as intermediate mesodermal progenitor (IMP) cells, is capable of unlimited expansion, lacks tumor formation potential, and, upon appropriate stimulation, readily acquires properties of a sub-population of kidney cells. Interestingly, IMP cells fail to differentiate into other mesodermally-derived tissues, including blood and heart, suggesting that these cells are restricted to an intermediate mesodermal fate.

No MeSH data available.


Related in: MedlinePlus

Assessment of renal potential of IMP cells.(A) Schematic of spinal cord (SC) co-culture assay to assess renal differentiation potential of IMP cells. IMP cells were differentiated as depicted in Figure 7A and incubated in liquid–air interface cultures with rat embryonic SC explants. (B) Immuno-fluorescence analysis of markers expressed in renal progenitors. 4 days after co-cultures were established, cells were fixed and stained for the indicated proteins (ECAD, SIX2 and SALL1) and for Lotus-tetragonolobus lectin (LTL). The dashed line indicates the boundary between human cells and the SC explant. Scale bar = 100 µm. (C) Undifferentiated hES cells failed to express SIX2 when co-cultured with embryonic rat SCs. Scale bar = 100 µm.DOI:http://dx.doi.org/10.7554/eLife.08413.021
© Copyright Policy
Related In: Results  -  Collection

License
getmorefigures.php?uid=PMC4631902&req=5

fig8: Assessment of renal potential of IMP cells.(A) Schematic of spinal cord (SC) co-culture assay to assess renal differentiation potential of IMP cells. IMP cells were differentiated as depicted in Figure 7A and incubated in liquid–air interface cultures with rat embryonic SC explants. (B) Immuno-fluorescence analysis of markers expressed in renal progenitors. 4 days after co-cultures were established, cells were fixed and stained for the indicated proteins (ECAD, SIX2 and SALL1) and for Lotus-tetragonolobus lectin (LTL). The dashed line indicates the boundary between human cells and the SC explant. Scale bar = 100 µm. (C) Undifferentiated hES cells failed to express SIX2 when co-cultured with embryonic rat SCs. Scale bar = 100 µm.DOI:http://dx.doi.org/10.7554/eLife.08413.021

Mentions: To further assess the ability of the IMP cells to generate cells with renal properties, we employed two rat explant assays that represent stringent measures of renal potential. In the first assay, we co-cultured IMP-derived MM cells with dissected embryonic rat spinal cords (SCs), a tissue that produces potent nephrogenic inductive signals (Figure 8A) (Kispert et al., 1998; Osafune et al., 2006; Gallegos et al., 2012). In this system, IMP-derived MM cells readily acquired expression of markers associated with renal cell types, including Lotus tetragonolobus lectin (LTL), CDH1, SALL1 and SIX2 (Figure 8B). In contrast, undifferentiated hES cells failed to express of SIX2 (Figure 8C), indicating that MM properties are required for efficient renal differentiation. Although IMP-derived MM cells expressed several markers associated with the renal lineage, they failed to generate tubule-like structures, including the nephron, suggesting that IMP cells differentiate effectively into a sub-population of kidney cells. These co-culture experiments demonstrate that IMP cells efficiently generate cell types with renal characteristics.10.7554/eLife.08413.021Figure 8.Assessment of renal potential of IMP cells.


Generation of an expandable intermediate mesoderm restricted progenitor cell line from human pluripotent stem cells.

Kumar N, Richter J, Cutts J, Bush KT, Trujillo C, Nigam SK, Gaasterland T, Brafman D, Willert K - Elife (2015)

Assessment of renal potential of IMP cells.(A) Schematic of spinal cord (SC) co-culture assay to assess renal differentiation potential of IMP cells. IMP cells were differentiated as depicted in Figure 7A and incubated in liquid–air interface cultures with rat embryonic SC explants. (B) Immuno-fluorescence analysis of markers expressed in renal progenitors. 4 days after co-cultures were established, cells were fixed and stained for the indicated proteins (ECAD, SIX2 and SALL1) and for Lotus-tetragonolobus lectin (LTL). The dashed line indicates the boundary between human cells and the SC explant. Scale bar = 100 µm. (C) Undifferentiated hES cells failed to express SIX2 when co-cultured with embryonic rat SCs. Scale bar = 100 µm.DOI:http://dx.doi.org/10.7554/eLife.08413.021
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4631902&req=5

fig8: Assessment of renal potential of IMP cells.(A) Schematic of spinal cord (SC) co-culture assay to assess renal differentiation potential of IMP cells. IMP cells were differentiated as depicted in Figure 7A and incubated in liquid–air interface cultures with rat embryonic SC explants. (B) Immuno-fluorescence analysis of markers expressed in renal progenitors. 4 days after co-cultures were established, cells were fixed and stained for the indicated proteins (ECAD, SIX2 and SALL1) and for Lotus-tetragonolobus lectin (LTL). The dashed line indicates the boundary between human cells and the SC explant. Scale bar = 100 µm. (C) Undifferentiated hES cells failed to express SIX2 when co-cultured with embryonic rat SCs. Scale bar = 100 µm.DOI:http://dx.doi.org/10.7554/eLife.08413.021
Mentions: To further assess the ability of the IMP cells to generate cells with renal properties, we employed two rat explant assays that represent stringent measures of renal potential. In the first assay, we co-cultured IMP-derived MM cells with dissected embryonic rat spinal cords (SCs), a tissue that produces potent nephrogenic inductive signals (Figure 8A) (Kispert et al., 1998; Osafune et al., 2006; Gallegos et al., 2012). In this system, IMP-derived MM cells readily acquired expression of markers associated with renal cell types, including Lotus tetragonolobus lectin (LTL), CDH1, SALL1 and SIX2 (Figure 8B). In contrast, undifferentiated hES cells failed to express of SIX2 (Figure 8C), indicating that MM properties are required for efficient renal differentiation. Although IMP-derived MM cells expressed several markers associated with the renal lineage, they failed to generate tubule-like structures, including the nephron, suggesting that IMP cells differentiate effectively into a sub-population of kidney cells. These co-culture experiments demonstrate that IMP cells efficiently generate cell types with renal characteristics.10.7554/eLife.08413.021Figure 8.Assessment of renal potential of IMP cells.

Bottom Line: One way to mitigate this risk is to develop expandable progenitor cell populations with restricted differentiation potential.Here, we used a cellular microarray technology to identify a defined and optimized culture condition that supports the derivation and propagation of a cell population with mesodermal properties.Interestingly, IMP cells fail to differentiate into other mesodermally-derived tissues, including blood and heart, suggesting that these cells are restricted to an intermediate mesodermal fate.

View Article: PubMed Central - PubMed

Affiliation: Department of Cellular and Molecular Medicine, University of California, San Diego, San Diego, United States.

ABSTRACT
The field of tissue engineering entered a new era with the development of human pluripotent stem cells (hPSCs), which are capable of unlimited expansion whilst retaining the potential to differentiate into all mature cell populations. However, these cells harbor significant risks, including tumor formation upon transplantation. One way to mitigate this risk is to develop expandable progenitor cell populations with restricted differentiation potential. Here, we used a cellular microarray technology to identify a defined and optimized culture condition that supports the derivation and propagation of a cell population with mesodermal properties. This cell population, referred to as intermediate mesodermal progenitor (IMP) cells, is capable of unlimited expansion, lacks tumor formation potential, and, upon appropriate stimulation, readily acquires properties of a sub-population of kidney cells. Interestingly, IMP cells fail to differentiate into other mesodermally-derived tissues, including blood and heart, suggesting that these cells are restricted to an intermediate mesodermal fate.

No MeSH data available.


Related in: MedlinePlus