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Cyclic ADP ribose is a novel regulator of intracellular Ca2+ oscillations in human bone marrow mesenchymal stem cells.

Tao R, Sun HY, Lau CP, Tse HF, Lee HC, Li GR - J. Cell. Mol. Med. (2011)

Bottom Line: Moreover, cADPR remarkably increased phosphorylated extracellular-signal-regulated kinases 1/2 (ERK1/2), but not Akt or p38 mitogen-activated protein kinase (MAPK).However, cADPR had no effect on adipogenesis or osteogenesis in human MSCs.This study delineates an alternate signalling pathway of cADPR that is distinct from its well-established role of serving as a Ca(2+) messenger for mobilizing the internal Ca(2+) stores.

View Article: PubMed Central - PubMed

Affiliation: Department of Medicine, Li Ka Shing Faculty of Medicine, The University of Hong Kong, Pokfulam, Hong Kong SAR, China.

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Characteristics of human MSCs. (A) Cells in culture on day 3 (left) and day 8 (right) from subculture from passage 4. (B) Adipogenic differentiation of human MSCs (passage 5) in control (left) and adipogenic supplement (right). Cells were stained with Oil red O. (C) Osteogenic differentiation of human MSCs (passage 5) in control (left) and osteogenic supplement (right). Cells were stained with Alizarin red S. (D) RT-PCR revealed the lineage-specific genes expression in cells from controls and adipogenic and osteogenic differentiation cultures. PPAR-γ: peroxisome proliferator-activated receptor gamma; FABP4: fatty acid binding protein 4; RUNX2: runt related transcription factor 2. (E) ‘Pseudo’-colour images show changes in fluorescence intensity (i.e. Ca2+i) in the cell at time-points a, b, c, d, as indicated in (F). (F) Spontaneous Ca2+i oscillations observed in a representative cell from passage 3. The pseudo-ratio δF/F0: δF/F0= (F − Fbase)/Fbase was applied to express Ca2+i level, where F is the measured fluorescence intensity of Fluo-3, Fbase is the lowest level of fluorescence intensity in the cell. (G) Incidence of spontaneous Ca2+i oscillations in human MSCs from passages 2 and 5.
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fig01: Characteristics of human MSCs. (A) Cells in culture on day 3 (left) and day 8 (right) from subculture from passage 4. (B) Adipogenic differentiation of human MSCs (passage 5) in control (left) and adipogenic supplement (right). Cells were stained with Oil red O. (C) Osteogenic differentiation of human MSCs (passage 5) in control (left) and osteogenic supplement (right). Cells were stained with Alizarin red S. (D) RT-PCR revealed the lineage-specific genes expression in cells from controls and adipogenic and osteogenic differentiation cultures. PPAR-γ: peroxisome proliferator-activated receptor gamma; FABP4: fatty acid binding protein 4; RUNX2: runt related transcription factor 2. (E) ‘Pseudo’-colour images show changes in fluorescence intensity (i.e. Ca2+i) in the cell at time-points a, b, c, d, as indicated in (F). (F) Spontaneous Ca2+i oscillations observed in a representative cell from passage 3. The pseudo-ratio δF/F0: δF/F0= (F − Fbase)/Fbase was applied to express Ca2+i level, where F is the measured fluorescence intensity of Fluo-3, Fbase is the lowest level of fluorescence intensity in the cell. (G) Incidence of spontaneous Ca2+i oscillations in human MSCs from passages 2 and 5.

Mentions: Human MSCs (passage 4) showed a fibroblast-like appearance at day 3 after sub-culturing and became confluence at day 8 (Fig. 1). Figure 1B and C display adipogenic and osteogenic differentiation in human MSCs at passage 5. The cells could be induced to differentiate efficiently into adipocytes (Fig. 1B) or osteocytes (Fig. 1C), which occurred, however, only after specific induction. RT-PCR (Fig. 1D) revealed the expression of lineage-specific genes after induction: PPAR-γ and fatty acid binding protein 4 for adipocytes, and runt related transcription factor 2 and osteocalcin for osteocytes [2, 31]. The PCR primers used for detection of these specific genes are shown in Table S1. These results indicate that human MSCs we studied here are maintained in the ‘stem’ state before induction.


Cyclic ADP ribose is a novel regulator of intracellular Ca2+ oscillations in human bone marrow mesenchymal stem cells.

Tao R, Sun HY, Lau CP, Tse HF, Lee HC, Li GR - J. Cell. Mol. Med. (2011)

Characteristics of human MSCs. (A) Cells in culture on day 3 (left) and day 8 (right) from subculture from passage 4. (B) Adipogenic differentiation of human MSCs (passage 5) in control (left) and adipogenic supplement (right). Cells were stained with Oil red O. (C) Osteogenic differentiation of human MSCs (passage 5) in control (left) and osteogenic supplement (right). Cells were stained with Alizarin red S. (D) RT-PCR revealed the lineage-specific genes expression in cells from controls and adipogenic and osteogenic differentiation cultures. PPAR-γ: peroxisome proliferator-activated receptor gamma; FABP4: fatty acid binding protein 4; RUNX2: runt related transcription factor 2. (E) ‘Pseudo’-colour images show changes in fluorescence intensity (i.e. Ca2+i) in the cell at time-points a, b, c, d, as indicated in (F). (F) Spontaneous Ca2+i oscillations observed in a representative cell from passage 3. The pseudo-ratio δF/F0: δF/F0= (F − Fbase)/Fbase was applied to express Ca2+i level, where F is the measured fluorescence intensity of Fluo-3, Fbase is the lowest level of fluorescence intensity in the cell. (G) Incidence of spontaneous Ca2+i oscillations in human MSCs from passages 2 and 5.
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Related In: Results  -  Collection

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fig01: Characteristics of human MSCs. (A) Cells in culture on day 3 (left) and day 8 (right) from subculture from passage 4. (B) Adipogenic differentiation of human MSCs (passage 5) in control (left) and adipogenic supplement (right). Cells were stained with Oil red O. (C) Osteogenic differentiation of human MSCs (passage 5) in control (left) and osteogenic supplement (right). Cells were stained with Alizarin red S. (D) RT-PCR revealed the lineage-specific genes expression in cells from controls and adipogenic and osteogenic differentiation cultures. PPAR-γ: peroxisome proliferator-activated receptor gamma; FABP4: fatty acid binding protein 4; RUNX2: runt related transcription factor 2. (E) ‘Pseudo’-colour images show changes in fluorescence intensity (i.e. Ca2+i) in the cell at time-points a, b, c, d, as indicated in (F). (F) Spontaneous Ca2+i oscillations observed in a representative cell from passage 3. The pseudo-ratio δF/F0: δF/F0= (F − Fbase)/Fbase was applied to express Ca2+i level, where F is the measured fluorescence intensity of Fluo-3, Fbase is the lowest level of fluorescence intensity in the cell. (G) Incidence of spontaneous Ca2+i oscillations in human MSCs from passages 2 and 5.
Mentions: Human MSCs (passage 4) showed a fibroblast-like appearance at day 3 after sub-culturing and became confluence at day 8 (Fig. 1). Figure 1B and C display adipogenic and osteogenic differentiation in human MSCs at passage 5. The cells could be induced to differentiate efficiently into adipocytes (Fig. 1B) or osteocytes (Fig. 1C), which occurred, however, only after specific induction. RT-PCR (Fig. 1D) revealed the expression of lineage-specific genes after induction: PPAR-γ and fatty acid binding protein 4 for adipocytes, and runt related transcription factor 2 and osteocalcin for osteocytes [2, 31]. The PCR primers used for detection of these specific genes are shown in Table S1. These results indicate that human MSCs we studied here are maintained in the ‘stem’ state before induction.

Bottom Line: Moreover, cADPR remarkably increased phosphorylated extracellular-signal-regulated kinases 1/2 (ERK1/2), but not Akt or p38 mitogen-activated protein kinase (MAPK).However, cADPR had no effect on adipogenesis or osteogenesis in human MSCs.This study delineates an alternate signalling pathway of cADPR that is distinct from its well-established role of serving as a Ca(2+) messenger for mobilizing the internal Ca(2+) stores.

View Article: PubMed Central - PubMed

Affiliation: Department of Medicine, Li Ka Shing Faculty of Medicine, The University of Hong Kong, Pokfulam, Hong Kong SAR, China.

Show MeSH
Related in: MedlinePlus