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Bone marrow-derived cells can acquire cardiac stem cells properties in damaged heart.

Barile L, Cerisoli F, Frati G, Gaetani R, Chimenti I, Forte E, Cassinelli L, Spinardi L, Altomare C, Kizana E, Giacomello A, Messina E, Ottolenghi S, Magli MC - J. Cell. Mol. Med. (2011)

Bottom Line: However, there is no direct evidence, so far, that BM cells can generate cardiac stem cells (CSCs).Following haematological reconstitution and MI, CSCs were cultured from cardiac explants to generate 'cardiospheres', a microtissue normally originating in vitro from CSCs.These were all green fluorescent (i.e. BM derived) and contained cells capable of initiating differentiation into cells expressing the cardiac marker Nkx2.5.

View Article: PubMed Central - PubMed

Affiliation: Institut Pasteur-Cenci Bolognetti Foundation, Department of Experimental Medicine, University of Rome La Sapienza, Rome, Italy.

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CS and CSDCs phenotypes. Confocal analysis of a CS, derived from the infarcted heart of a lethally irradiated mouse transplanted with marrow cells of Kit/GFP transgenic mice (A) Merged image showing co-expression in some cells (arrows) of donor cell-derived GFP (green) and Nkx2.5 (an immature cardiac cell-specific transcription factor) (red); nuclei are stained blue by Hoechst dye. The figure is an average of 13 z-axis confocal sections. Single channel fluorescence intensity of some cells within the sphere is represented by the plots, depicting the fluorescence of cells traced by the white line. Co-localization is evident for the cells indicated as cell 1 and 2 whereas the third nucleus is expressing neither GFP nor Nkx2.5. (B) Single cell derived from CS dissociation (from a different CS), demonstrating nuclear co-localization of GFP and Nkx2.5. (C) RT-PCR of GFP, Nkx2.5 and cardiac actin from cells expanded on fibronectin from a single clonogenic CS, and later analysed as a monolayer on fibronectin (left) or as CSs (on polylisine).
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fig03: CS and CSDCs phenotypes. Confocal analysis of a CS, derived from the infarcted heart of a lethally irradiated mouse transplanted with marrow cells of Kit/GFP transgenic mice (A) Merged image showing co-expression in some cells (arrows) of donor cell-derived GFP (green) and Nkx2.5 (an immature cardiac cell-specific transcription factor) (red); nuclei are stained blue by Hoechst dye. The figure is an average of 13 z-axis confocal sections. Single channel fluorescence intensity of some cells within the sphere is represented by the plots, depicting the fluorescence of cells traced by the white line. Co-localization is evident for the cells indicated as cell 1 and 2 whereas the third nucleus is expressing neither GFP nor Nkx2.5. (B) Single cell derived from CS dissociation (from a different CS), demonstrating nuclear co-localization of GFP and Nkx2.5. (C) RT-PCR of GFP, Nkx2.5 and cardiac actin from cells expanded on fibronectin from a single clonogenic CS, and later analysed as a monolayer on fibronectin (left) or as CSs (on polylisine).

Mentions: CSs grow by expansion of a central core of immature cells, which initiate differentiation in the course of their displacement to the periphery of the sphere; this results in the appearance of a small number of cells positive for Nkx2.5, a transcription factor representing an early cardiac marker, and occasionally Troponin I, a latter cytoplasmic marker [1]. A very low level of in vitro differentiation of a type of Kit+ CSC was also reported by Anversa’s group [6]. A representative pool of about 120 CSs, derived from six different transplanted mice, were collected and individually stained with antibodies against Nkx2.5 or Troponin I to evaluate both antibody staining and GFP fluorescence. Confocal analysis shows cells that express GFP together with Nkx2.5 (Figs 3A, B and S2). The percentage of GFP+ cells co-expressing Nkx2.5 is rather variable, ranging between 5% and 50% and depends on the developmental stage of the CSs. Conversely, a very low level of GFP, if any, is stained by anti-TnI-antibody (data not shown). This suggests that cells, which occasionally commit to cardiomyocyte differentiation (i.e. are Nkx2.5+), progressively extinguish Kit/GFP expression, while up-regulating cardiac-specific genes. Interestingly, when a GFP+ CS is dissociated and the resulting cells are replated in fibronectin, the kit/GFP gene is substantially down-regulated; however, if the cells are again grown on polylisine, GFP expression is reactivated (Fig. 3C). This is in agreement with previous findings by Messina et al. [1]. Thus, BM-derived cells present in the CS may express cardiac markers.


Bone marrow-derived cells can acquire cardiac stem cells properties in damaged heart.

Barile L, Cerisoli F, Frati G, Gaetani R, Chimenti I, Forte E, Cassinelli L, Spinardi L, Altomare C, Kizana E, Giacomello A, Messina E, Ottolenghi S, Magli MC - J. Cell. Mol. Med. (2011)

CS and CSDCs phenotypes. Confocal analysis of a CS, derived from the infarcted heart of a lethally irradiated mouse transplanted with marrow cells of Kit/GFP transgenic mice (A) Merged image showing co-expression in some cells (arrows) of donor cell-derived GFP (green) and Nkx2.5 (an immature cardiac cell-specific transcription factor) (red); nuclei are stained blue by Hoechst dye. The figure is an average of 13 z-axis confocal sections. Single channel fluorescence intensity of some cells within the sphere is represented by the plots, depicting the fluorescence of cells traced by the white line. Co-localization is evident for the cells indicated as cell 1 and 2 whereas the third nucleus is expressing neither GFP nor Nkx2.5. (B) Single cell derived from CS dissociation (from a different CS), demonstrating nuclear co-localization of GFP and Nkx2.5. (C) RT-PCR of GFP, Nkx2.5 and cardiac actin from cells expanded on fibronectin from a single clonogenic CS, and later analysed as a monolayer on fibronectin (left) or as CSs (on polylisine).
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Related In: Results  -  Collection

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fig03: CS and CSDCs phenotypes. Confocal analysis of a CS, derived from the infarcted heart of a lethally irradiated mouse transplanted with marrow cells of Kit/GFP transgenic mice (A) Merged image showing co-expression in some cells (arrows) of donor cell-derived GFP (green) and Nkx2.5 (an immature cardiac cell-specific transcription factor) (red); nuclei are stained blue by Hoechst dye. The figure is an average of 13 z-axis confocal sections. Single channel fluorescence intensity of some cells within the sphere is represented by the plots, depicting the fluorescence of cells traced by the white line. Co-localization is evident for the cells indicated as cell 1 and 2 whereas the third nucleus is expressing neither GFP nor Nkx2.5. (B) Single cell derived from CS dissociation (from a different CS), demonstrating nuclear co-localization of GFP and Nkx2.5. (C) RT-PCR of GFP, Nkx2.5 and cardiac actin from cells expanded on fibronectin from a single clonogenic CS, and later analysed as a monolayer on fibronectin (left) or as CSs (on polylisine).
Mentions: CSs grow by expansion of a central core of immature cells, which initiate differentiation in the course of their displacement to the periphery of the sphere; this results in the appearance of a small number of cells positive for Nkx2.5, a transcription factor representing an early cardiac marker, and occasionally Troponin I, a latter cytoplasmic marker [1]. A very low level of in vitro differentiation of a type of Kit+ CSC was also reported by Anversa’s group [6]. A representative pool of about 120 CSs, derived from six different transplanted mice, were collected and individually stained with antibodies against Nkx2.5 or Troponin I to evaluate both antibody staining and GFP fluorescence. Confocal analysis shows cells that express GFP together with Nkx2.5 (Figs 3A, B and S2). The percentage of GFP+ cells co-expressing Nkx2.5 is rather variable, ranging between 5% and 50% and depends on the developmental stage of the CSs. Conversely, a very low level of GFP, if any, is stained by anti-TnI-antibody (data not shown). This suggests that cells, which occasionally commit to cardiomyocyte differentiation (i.e. are Nkx2.5+), progressively extinguish Kit/GFP expression, while up-regulating cardiac-specific genes. Interestingly, when a GFP+ CS is dissociated and the resulting cells are replated in fibronectin, the kit/GFP gene is substantially down-regulated; however, if the cells are again grown on polylisine, GFP expression is reactivated (Fig. 3C). This is in agreement with previous findings by Messina et al. [1]. Thus, BM-derived cells present in the CS may express cardiac markers.

Bottom Line: However, there is no direct evidence, so far, that BM cells can generate cardiac stem cells (CSCs).Following haematological reconstitution and MI, CSCs were cultured from cardiac explants to generate 'cardiospheres', a microtissue normally originating in vitro from CSCs.These were all green fluorescent (i.e. BM derived) and contained cells capable of initiating differentiation into cells expressing the cardiac marker Nkx2.5.

View Article: PubMed Central - PubMed

Affiliation: Institut Pasteur-Cenci Bolognetti Foundation, Department of Experimental Medicine, University of Rome La Sapienza, Rome, Italy.

Show MeSH
Related in: MedlinePlus