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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

Molecular characteristics of CSCs/CPCs during chasing and post-injury(A)–(C) Show staining of cardiac tissues in 2 months chasing. (A) indicates staining of BrdU and c-Kit, the 2 fine arrows point to double positive cells, and the asterisk indicates BrdU+/c-Kit− cells (asterisk); however, the thick arrow points to BrdU−/c-Kit+ cells (arrow). (B) Indicates staining of BrdU and Sca-1, a fine arrow points to double positive cells, and the asterisk indicates BrdU+ and Sca-1− cells; however, the thick arrow points to BrdU−/Sca-1+ cells (arrow). (C) Indicates staining of BrdU and N-cad, the arrowhead indicates BrdU+/N-cad+ cells adhering to BrdU−/N-cad+ spindle cells (arrow), the low right corner in (C) are magnification of rectangular region, and the hollow arrow indicate N-cad expression between the BrdU+ cells and BrdU− cells. (D and E) Show staining of cardiac tissues at day 4 post-injury; (D) indicates staining of BrdU and c-Kit. The 2 fine arrow point to double positive cells; however, the thick arrow points to BrdU−/c-Kit+ cells (arrow). (E) Indicates staining of BrdU and Sca-1, fine arrow point to double positive cells, however, the thick arrow points to BrdU−/Sca-1+ cells (arrow). Nuclei were staining with DAPI. (F) Shows statistical of double positive cells. *P<0.01 show most significance and **P>0.05 shows no significance. (G) Shows a dynamic model of CSCs/CPCs regeneration post-injury. The d-CSCs/CPCs pool (d-LRC-CSCs/CPCs), circulate only a few times over the life span of the mouse in a homoeostatic situation, but is activated (a-LRC-CSCs/CPCs) and participated in replenishment of the cardiac tissues post-injury (scale bars: A, B, D and E = 20 μm; C = 10 μm).
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Figure 4: Molecular characteristics of CSCs/CPCs during chasing and post-injury(A)–(C) Show staining of cardiac tissues in 2 months chasing. (A) indicates staining of BrdU and c-Kit, the 2 fine arrows point to double positive cells, and the asterisk indicates BrdU+/c-Kit− cells (asterisk); however, the thick arrow points to BrdU−/c-Kit+ cells (arrow). (B) Indicates staining of BrdU and Sca-1, a fine arrow points to double positive cells, and the asterisk indicates BrdU+ and Sca-1− cells; however, the thick arrow points to BrdU−/Sca-1+ cells (arrow). (C) Indicates staining of BrdU and N-cad, the arrowhead indicates BrdU+/N-cad+ cells adhering to BrdU−/N-cad+ spindle cells (arrow), the low right corner in (C) are magnification of rectangular region, and the hollow arrow indicate N-cad expression between the BrdU+ cells and BrdU− cells. (D and E) Show staining of cardiac tissues at day 4 post-injury; (D) indicates staining of BrdU and c-Kit. The 2 fine arrow point to double positive cells; however, the thick arrow points to BrdU−/c-Kit+ cells (arrow). (E) Indicates staining of BrdU and Sca-1, fine arrow point to double positive cells, however, the thick arrow points to BrdU−/Sca-1+ cells (arrow). Nuclei were staining with DAPI. (F) Shows statistical of double positive cells. *P<0.01 show most significance and **P>0.05 shows no significance. (G) Shows a dynamic model of CSCs/CPCs regeneration post-injury. The d-CSCs/CPCs pool (d-LRC-CSCs/CPCs), circulate only a few times over the life span of the mouse in a homoeostatic situation, but is activated (a-LRC-CSCs/CPCs) and participated in replenishment of the cardiac tissues post-injury (scale bars: A, B, D and E = 20 μm; C = 10 μm).

Mentions: Two weeks after the initial pulse (Figures 1B and 1C), the labelled cells were mainly myocardium cells with various size and oval-like cells located in the vicinity of the vascular endothelia region, and most of the labelled cells were brightly stained with BrdU. These cells were located primarily in the apex (Figure 1BI), interventricular septum (Figure 1BII), left ventricle (Figure 1BIII), cardiac valves (Figure 1BIV) and vascular endothelial region (Figure 1C). Moreover, they appeared in clusters. After 3 months of chasing, bright labelled BrdU (BrdU bright) cells were mainly myocardium cells, located in the cardiac valves (Figure 1D) and vascular endothelial region (Figure 1E). The labelled cells still appeared in cluster; however, they showed a trend to a more scattered distribution after prolonged chasing. However, BrdU positive cells were not found between the myocardial layers. Interestingly, several ‘BrdU bright’ cells appeared in the vicinity of the vascular region with varied embedding, and their nuclei were oval-shaped, which suggested the existence of vascular endothelium progenitor cells with the property of retaining BrdU label. In addition, a part of the LRCs also expressed the Sca c-Kit or Sca-1 (Figures 4A and 4B).


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

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

Molecular characteristics of CSCs/CPCs during chasing and post-injury(A)–(C) Show staining of cardiac tissues in 2 months chasing. (A) indicates staining of BrdU and c-Kit, the 2 fine arrows point to double positive cells, and the asterisk indicates BrdU+/c-Kit− cells (asterisk); however, the thick arrow points to BrdU−/c-Kit+ cells (arrow). (B) Indicates staining of BrdU and Sca-1, a fine arrow points to double positive cells, and the asterisk indicates BrdU+ and Sca-1− cells; however, the thick arrow points to BrdU−/Sca-1+ cells (arrow). (C) Indicates staining of BrdU and N-cad, the arrowhead indicates BrdU+/N-cad+ cells adhering to BrdU−/N-cad+ spindle cells (arrow), the low right corner in (C) are magnification of rectangular region, and the hollow arrow indicate N-cad expression between the BrdU+ cells and BrdU− cells. (D and E) Show staining of cardiac tissues at day 4 post-injury; (D) indicates staining of BrdU and c-Kit. The 2 fine arrow point to double positive cells; however, the thick arrow points to BrdU−/c-Kit+ cells (arrow). (E) Indicates staining of BrdU and Sca-1, fine arrow point to double positive cells, however, the thick arrow points to BrdU−/Sca-1+ cells (arrow). Nuclei were staining with DAPI. (F) Shows statistical of double positive cells. *P<0.01 show most significance and **P>0.05 shows no significance. (G) Shows a dynamic model of CSCs/CPCs regeneration post-injury. The d-CSCs/CPCs pool (d-LRC-CSCs/CPCs), circulate only a few times over the life span of the mouse in a homoeostatic situation, but is activated (a-LRC-CSCs/CPCs) and participated in replenishment of the cardiac tissues post-injury (scale bars: A, B, D and E = 20 μm; C = 10 μm).
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Related In: Results  -  Collection

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Figure 4: Molecular characteristics of CSCs/CPCs during chasing and post-injury(A)–(C) Show staining of cardiac tissues in 2 months chasing. (A) indicates staining of BrdU and c-Kit, the 2 fine arrows point to double positive cells, and the asterisk indicates BrdU+/c-Kit− cells (asterisk); however, the thick arrow points to BrdU−/c-Kit+ cells (arrow). (B) Indicates staining of BrdU and Sca-1, a fine arrow points to double positive cells, and the asterisk indicates BrdU+ and Sca-1− cells; however, the thick arrow points to BrdU−/Sca-1+ cells (arrow). (C) Indicates staining of BrdU and N-cad, the arrowhead indicates BrdU+/N-cad+ cells adhering to BrdU−/N-cad+ spindle cells (arrow), the low right corner in (C) are magnification of rectangular region, and the hollow arrow indicate N-cad expression between the BrdU+ cells and BrdU− cells. (D and E) Show staining of cardiac tissues at day 4 post-injury; (D) indicates staining of BrdU and c-Kit. The 2 fine arrow point to double positive cells; however, the thick arrow points to BrdU−/c-Kit+ cells (arrow). (E) Indicates staining of BrdU and Sca-1, fine arrow point to double positive cells, however, the thick arrow points to BrdU−/Sca-1+ cells (arrow). Nuclei were staining with DAPI. (F) Shows statistical of double positive cells. *P<0.01 show most significance and **P>0.05 shows no significance. (G) Shows a dynamic model of CSCs/CPCs regeneration post-injury. The d-CSCs/CPCs pool (d-LRC-CSCs/CPCs), circulate only a few times over the life span of the mouse in a homoeostatic situation, but is activated (a-LRC-CSCs/CPCs) and participated in replenishment of the cardiac tissues post-injury (scale bars: A, B, D and E = 20 μm; C = 10 μm).
Mentions: Two weeks after the initial pulse (Figures 1B and 1C), the labelled cells were mainly myocardium cells with various size and oval-like cells located in the vicinity of the vascular endothelia region, and most of the labelled cells were brightly stained with BrdU. These cells were located primarily in the apex (Figure 1BI), interventricular septum (Figure 1BII), left ventricle (Figure 1BIII), cardiac valves (Figure 1BIV) and vascular endothelial region (Figure 1C). Moreover, they appeared in clusters. After 3 months of chasing, bright labelled BrdU (BrdU bright) cells were mainly myocardium cells, located in the cardiac valves (Figure 1D) and vascular endothelial region (Figure 1E). The labelled cells still appeared in cluster; however, they showed a trend to a more scattered distribution after prolonged chasing. However, BrdU positive cells were not found between the myocardial layers. Interestingly, several ‘BrdU bright’ cells appeared in the vicinity of the vascular region with varied embedding, and their nuclei were oval-shaped, which suggested the existence of vascular endothelium progenitor cells with the property of retaining BrdU label. In addition, a part of the LRCs also expressed the Sca c-Kit or Sca-1 (Figures 4A and 4B).

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