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Identification of functional tissue-resident cardiac stem/progenitor cells in adult mouse.

Lu L, Li F, Lu J - Cell Biol Int Rep (2010) (2012)

Bottom Line: LRCs that definitively exist in the heart tissues of adult mice, and some LRCs express the stem cell marker, Sca-1 or c-Kit, and are located primarily in the myocardium and vascular endothelial regions.Moreover, the number of LRCs remains nearly constant during the lifespan of the mouse.A small percentage of the CSCs/CPCs express Sca-1 or c-Kit.

View Article: PubMed Central - PubMed

Affiliation: Department of Bimolecular Engineering, Tohoku University, Sendai 9808579, Miyagi, Japan ; †Institute for Regeneration Medicine, Changzhou Tenraid Biotech Co., Ltd, Changzhou, Jiangsu, 213022, People's Republic of China ; §School of Pharmaceutical Engineering and Life Science, Changzhou University, Changzhou, Jiangsu 213164, People's Republic of China.

ABSTRACT
In most somatic tissues, ASCs (adult stem cells) are crucial for the maintenance of tissue homoeostasis under normal physiological state and recovery from injury. LRC (label retaining cell) assay is a well-known method of identifying possible somatic stem/progenitor cells and their location both in situ and in vivo. BrdU (bromodeoxyuridine) was used here to tag the possible CSCs (cardiac stem cells)/CPCs (cardiac progenitor cells) in newborn pups, followed by a trace period of up to 24 months. In addition, we have used our newly developed 'KAL' method to rapidly Kill proliferating cells in adult heart tissues, then, Activate and Label the surviving CSCs/CPCs. LRCs that definitively exist in the heart tissues of adult mice, and some LRCs express the stem cell marker, Sca-1 or c-Kit, and are located primarily in the myocardium and vascular endothelial regions. Moreover, the number of LRCs remains nearly constant during the lifespan of the mouse. After injury induced by 5-fluorouracil, the proliferating cells were almost completely cleared on day 3, and the activated CSCs/CPCs retained their BrdU label after regeneration was complete. A small percentage of the CSCs/CPCs express Sca-1 or c-Kit. Furthermore, the LRC method together with KAL may be used to identify and locate possible CSCs/CPCs, which has potential clinical application.

No MeSH data available.


Related in: MedlinePlus

Immunohistostaining with BrdU and DAPI of the control and the mice at various days post 5-Fu exposure(A) Shows staining of a control heart section. (B) Shows staining on day 3. (C) Shows staining on day 4. (D) Shows staining on day 5. (E) Shows staining on day 7. (F) Shows the statistical analysis of BrdU positive cells at each time point (n = 3 mice). Nuclei were stained with DAPI (blue), and the asterisks indicate ‘BrdU bright’ cells. *P<0.01 (control versus day 3 post-injury) and **P<0.01 (control versus day 4 post-injury) indicates most significant differences (scale bar = 20 μm).
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Figure 2: Immunohistostaining with BrdU and DAPI of the control and the mice at various days post 5-Fu exposure(A) Shows staining of a control heart section. (B) Shows staining on day 3. (C) Shows staining on day 4. (D) Shows staining on day 5. (E) Shows staining on day 7. (F) Shows the statistical analysis of BrdU positive cells at each time point (n = 3 mice). Nuclei were stained with DAPI (blue), and the asterisks indicate ‘BrdU bright’ cells. *P<0.01 (control versus day 3 post-injury) and **P<0.01 (control versus day 4 post-injury) indicates most significant differences (scale bar = 20 μm).

Mentions: We determined whether d-CSCs/CPCs can be activated and regenerate injured heart tissue, and also the kinetics of the activated CSCs/CPCs at various times post-injury. BrdU was administrated on days 3, 4, 5 and 7 to chase the activated CSCs/CPCs post-injury. The mice were killed 3 h after BrdU injection. After immunohistostaining of BrdU and DAPI, a few of the BrdU positive cells appeared in the myocardium in the control group (Figure 2A). On day 3 (Figure 2B), BrdU positive cells barely appeared in the heart tissues; however, on day 4, BrdU positive cells with variable shapes increased sharply, which suggested CSCs/CPCs were activated significantly from stem cell pool. After BrdU had been administrated on day 5 and day 7 post-injury, only a few BrdU positive cells appeared in heart tissues, and the number of LRCs gradually returned to the baseline level (Figures 2D and 2E), which suggested that the tissue-resident CSCs/CPCs were controlling the regeneration of the injured heart, and the activated-CSCs/CPCs returned to dormancy again after regeneration had been completed.


Identification of functional tissue-resident cardiac stem/progenitor cells in adult mouse.

Lu L, Li F, Lu J - Cell Biol Int Rep (2010) (2012)

Immunohistostaining with BrdU and DAPI of the control and the mice at various days post 5-Fu exposure(A) Shows staining of a control heart section. (B) Shows staining on day 3. (C) Shows staining on day 4. (D) Shows staining on day 5. (E) Shows staining on day 7. (F) Shows the statistical analysis of BrdU positive cells at each time point (n = 3 mice). Nuclei were stained with DAPI (blue), and the asterisks indicate ‘BrdU bright’ cells. *P<0.01 (control versus day 3 post-injury) and **P<0.01 (control versus day 4 post-injury) indicates most significant differences (scale bar = 20 μm).
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC3475443&req=5

Figure 2: Immunohistostaining with BrdU and DAPI of the control and the mice at various days post 5-Fu exposure(A) Shows staining of a control heart section. (B) Shows staining on day 3. (C) Shows staining on day 4. (D) Shows staining on day 5. (E) Shows staining on day 7. (F) Shows the statistical analysis of BrdU positive cells at each time point (n = 3 mice). Nuclei were stained with DAPI (blue), and the asterisks indicate ‘BrdU bright’ cells. *P<0.01 (control versus day 3 post-injury) and **P<0.01 (control versus day 4 post-injury) indicates most significant differences (scale bar = 20 μm).
Mentions: We determined whether d-CSCs/CPCs can be activated and regenerate injured heart tissue, and also the kinetics of the activated CSCs/CPCs at various times post-injury. BrdU was administrated on days 3, 4, 5 and 7 to chase the activated CSCs/CPCs post-injury. The mice were killed 3 h after BrdU injection. After immunohistostaining of BrdU and DAPI, a few of the BrdU positive cells appeared in the myocardium in the control group (Figure 2A). On day 3 (Figure 2B), BrdU positive cells barely appeared in the heart tissues; however, on day 4, BrdU positive cells with variable shapes increased sharply, which suggested CSCs/CPCs were activated significantly from stem cell pool. After BrdU had been administrated on day 5 and day 7 post-injury, only a few BrdU positive cells appeared in heart tissues, and the number of LRCs gradually returned to the baseline level (Figures 2D and 2E), which suggested that the tissue-resident CSCs/CPCs were controlling the regeneration of the injured heart, and the activated-CSCs/CPCs returned to dormancy again after regeneration had been completed.

Bottom Line: LRCs that definitively exist in the heart tissues of adult mice, and some LRCs express the stem cell marker, Sca-1 or c-Kit, and are located primarily in the myocardium and vascular endothelial regions.Moreover, the number of LRCs remains nearly constant during the lifespan of the mouse.A small percentage of the CSCs/CPCs express Sca-1 or c-Kit.

View Article: PubMed Central - PubMed

Affiliation: Department of Bimolecular Engineering, Tohoku University, Sendai 9808579, Miyagi, Japan ; †Institute for Regeneration Medicine, Changzhou Tenraid Biotech Co., Ltd, Changzhou, Jiangsu, 213022, People's Republic of China ; §School of Pharmaceutical Engineering and Life Science, Changzhou University, Changzhou, Jiangsu 213164, People's Republic of China.

ABSTRACT
In most somatic tissues, ASCs (adult stem cells) are crucial for the maintenance of tissue homoeostasis under normal physiological state and recovery from injury. LRC (label retaining cell) assay is a well-known method of identifying possible somatic stem/progenitor cells and their location both in situ and in vivo. BrdU (bromodeoxyuridine) was used here to tag the possible CSCs (cardiac stem cells)/CPCs (cardiac progenitor cells) in newborn pups, followed by a trace period of up to 24 months. In addition, we have used our newly developed 'KAL' method to rapidly Kill proliferating cells in adult heart tissues, then, Activate and Label the surviving CSCs/CPCs. LRCs that definitively exist in the heart tissues of adult mice, and some LRCs express the stem cell marker, Sca-1 or c-Kit, and are located primarily in the myocardium and vascular endothelial regions. Moreover, the number of LRCs remains nearly constant during the lifespan of the mouse. After injury induced by 5-fluorouracil, the proliferating cells were almost completely cleared on day 3, and the activated CSCs/CPCs retained their BrdU label after regeneration was complete. A small percentage of the CSCs/CPCs express Sca-1 or c-Kit. Furthermore, the LRC method together with KAL may be used to identify and locate possible CSCs/CPCs, which has potential clinical application.

No MeSH data available.


Related in: MedlinePlus