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Direct reprogramming of human bone marrow stromal cells into functional renal cells using cell-free extracts.

Papadimou E, Morigi M, Iatropoulos P, Xinaris C, Tomasoni S, Benedetti V, Longaretti L, Rota C, Todeschini M, Rizzo P, Introna M, Grazia de Simoni M, Remuzzi G, Goligorsky MS, Benigni A - Stem Cell Reports (2015)

Bottom Line: Transmission electron microscopy revealed the presence of brush border microvilli and tight intercellular contacts.RNA sequencing showed tubular epithelial transcript abundance and revealed the upregulation of components of the EGFR pathway.Thus, reprogrammed BMSCs are a promising cell resource for future cell therapy.

View Article: PubMed Central - PubMed

Affiliation: IRCCS-Istituto di Ricerche Farmacologiche "Mario Negri," Centro Anna Maria Astori, Science and Technology Park Kilometro Rosso, 24126 Bergamo, Italy. Electronic address: evangelia.papadimou@marionegri.it.

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Functional Properties of CL17 Cells In Vitro(A) Transepithelial resistance (TER) measurement of CL17, HK2, and BMSCs. Three independent experiments were done in duplicate. Data are expressed as mean ± SD; ∗p < 0.01 versus BMSCs.(B) BSA binding and uptake in CL17, HK2, and BMSCs. Cells were exposed to 50 μg/ml BSA-FITC alone or in the presence of excess cold BSA (5 mg/ml) for 15 min at 4°C for binding experiments (left) and for 90 min at 37°C for uptake experiments (right). Images are representative of three experiments. Scale bars, 50 μm.
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fig4: Functional Properties of CL17 Cells In Vitro(A) Transepithelial resistance (TER) measurement of CL17, HK2, and BMSCs. Three independent experiments were done in duplicate. Data are expressed as mean ± SD; ∗p < 0.01 versus BMSCs.(B) BSA binding and uptake in CL17, HK2, and BMSCs. Cells were exposed to 50 μg/ml BSA-FITC alone or in the presence of excess cold BSA (5 mg/ml) for 15 min at 4°C for binding experiments (left) and for 90 min at 37°C for uptake experiments (right). Images are representative of three experiments. Scale bars, 50 μm.

Mentions: Having observed the formation of nascent intercellular contacts, we next inquired whether CL17 cells in addition to morphological and cell-lineage markers had acquired physiological properties similar to the epithelial phenotype. Testing the electrical resistance of BMSC monolayers showed trivial values of 34 ± 10 Ohm/cm2. In contrast, monolayers of CL17 cells developed electrical resistance similar to that observed in HK2 cells (80 ± 10 versus 81 ± 12 Ohm/cm2) (Figure 4A), both statistically significantly different from the transepithelial resistance (TER) assessed in BMSCs.


Direct reprogramming of human bone marrow stromal cells into functional renal cells using cell-free extracts.

Papadimou E, Morigi M, Iatropoulos P, Xinaris C, Tomasoni S, Benedetti V, Longaretti L, Rota C, Todeschini M, Rizzo P, Introna M, Grazia de Simoni M, Remuzzi G, Goligorsky MS, Benigni A - Stem Cell Reports (2015)

Functional Properties of CL17 Cells In Vitro(A) Transepithelial resistance (TER) measurement of CL17, HK2, and BMSCs. Three independent experiments were done in duplicate. Data are expressed as mean ± SD; ∗p < 0.01 versus BMSCs.(B) BSA binding and uptake in CL17, HK2, and BMSCs. Cells were exposed to 50 μg/ml BSA-FITC alone or in the presence of excess cold BSA (5 mg/ml) for 15 min at 4°C for binding experiments (left) and for 90 min at 37°C for uptake experiments (right). Images are representative of three experiments. Scale bars, 50 μm.
© Copyright Policy - CC BY-NC-ND
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4400646&req=5

fig4: Functional Properties of CL17 Cells In Vitro(A) Transepithelial resistance (TER) measurement of CL17, HK2, and BMSCs. Three independent experiments were done in duplicate. Data are expressed as mean ± SD; ∗p < 0.01 versus BMSCs.(B) BSA binding and uptake in CL17, HK2, and BMSCs. Cells were exposed to 50 μg/ml BSA-FITC alone or in the presence of excess cold BSA (5 mg/ml) for 15 min at 4°C for binding experiments (left) and for 90 min at 37°C for uptake experiments (right). Images are representative of three experiments. Scale bars, 50 μm.
Mentions: Having observed the formation of nascent intercellular contacts, we next inquired whether CL17 cells in addition to morphological and cell-lineage markers had acquired physiological properties similar to the epithelial phenotype. Testing the electrical resistance of BMSC monolayers showed trivial values of 34 ± 10 Ohm/cm2. In contrast, monolayers of CL17 cells developed electrical resistance similar to that observed in HK2 cells (80 ± 10 versus 81 ± 12 Ohm/cm2) (Figure 4A), both statistically significantly different from the transepithelial resistance (TER) assessed in BMSCs.

Bottom Line: Transmission electron microscopy revealed the presence of brush border microvilli and tight intercellular contacts.RNA sequencing showed tubular epithelial transcript abundance and revealed the upregulation of components of the EGFR pathway.Thus, reprogrammed BMSCs are a promising cell resource for future cell therapy.

View Article: PubMed Central - PubMed

Affiliation: IRCCS-Istituto di Ricerche Farmacologiche "Mario Negri," Centro Anna Maria Astori, Science and Technology Park Kilometro Rosso, 24126 Bergamo, Italy. Electronic address: evangelia.papadimou@marionegri.it.

Show MeSH
Related in: MedlinePlus