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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

Existence and turnover of LRCs in cardiac tissues of normal adult mice(A) Shows schematic outlines of the BrdU pulse chase. w: week, m: month. (BI–IV) Show distinct locations of LRCs in the cardiac tissues after a 2 weeks chase, and the asterisk in I–IV indicates BrdU positive cells (asterisk), and the photo in the top left corner of IV contains white light. (C) Shows immunohistostaining results of BrdU (green) and CD31 (red) for cardiac tissues, and arrowhead indicates the location of LRCs after a 2 weeks chase. (D) Shows LRCs are mainly myocardium cells after a 3 months chase, and the asterisk in I–III shows BrdU (red) positive cells. (E) Shows LRCs located in the vicinity of vascular endothelial region with varied embedding after 3 months chase, and 6 asterisks indicate BrdU positive cells. (F) Shows identified LRCs in cardiac tissues after 24 months chase, and asterisk indicates BrdU positive myocardium cells, however, an arrow indicates ‘BrdU light’ myocardium cells. (G) Shows statistical analysis of BrdU label-retaining cells. The black boxes represent the mean values of LRCs (n = 2 mice at each time point). The error bars represent standard derivations. P = 0.026<0.05 shows there was significant difference (2 weeks versus 6 months), however, P = 0.103>0.05 indicates there was no significant difference (6 months versus 24 months). (H) Shows a model of LRCs in heart after birth, a part of activated-LRCs turn into d-LRCs. Scale bars: BI, BIII, C, E and F = 20 μm; DI–DIII = 30 μm; BII, BIV =  40 μm.
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Figure 1: Existence and turnover of LRCs in cardiac tissues of normal adult mice(A) Shows schematic outlines of the BrdU pulse chase. w: week, m: month. (BI–IV) Show distinct locations of LRCs in the cardiac tissues after a 2 weeks chase, and the asterisk in I–IV indicates BrdU positive cells (asterisk), and the photo in the top left corner of IV contains white light. (C) Shows immunohistostaining results of BrdU (green) and CD31 (red) for cardiac tissues, and arrowhead indicates the location of LRCs after a 2 weeks chase. (D) Shows LRCs are mainly myocardium cells after a 3 months chase, and the asterisk in I–III shows BrdU (red) positive cells. (E) Shows LRCs located in the vicinity of vascular endothelial region with varied embedding after 3 months chase, and 6 asterisks indicate BrdU positive cells. (F) Shows identified LRCs in cardiac tissues after 24 months chase, and asterisk indicates BrdU positive myocardium cells, however, an arrow indicates ‘BrdU light’ myocardium cells. (G) Shows statistical analysis of BrdU label-retaining cells. The black boxes represent the mean values of LRCs (n = 2 mice at each time point). The error bars represent standard derivations. P = 0.026<0.05 shows there was significant difference (2 weeks versus 6 months), however, P = 0.103>0.05 indicates there was no significant difference (6 months versus 24 months). (H) Shows a model of LRCs in heart after birth, a part of activated-LRCs turn into d-LRCs. Scale bars: BI, BIII, C, E and F = 20 μm; DI–DIII = 30 μm; BII, BIV =  40 μm.

Mentions: Newborn mice (7 days old) were injected subcutaneously with BrdU (Sigma, pH 7.4) twice a day for 3–5 consecutive days (50–100 mg/kg per injection for each mouse). The labelled mice were killed at 2 weeks (n = 2 mice), 3 months (n = 2 mice), 6 months (n = 2 mice) and 24 months (n = 2 mice) (Figure 1A) to trace and locate the LRCs throughout their lifespan. The traced time was calculated from the initial administration of BrdU. According to Urbanek et al. (2006), cells with BrdU label can be divided into ‘BrdU bright’ cells and ‘BrdU dim’ cells.


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

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

Existence and turnover of LRCs in cardiac tissues of normal adult mice(A) Shows schematic outlines of the BrdU pulse chase. w: week, m: month. (BI–IV) Show distinct locations of LRCs in the cardiac tissues after a 2 weeks chase, and the asterisk in I–IV indicates BrdU positive cells (asterisk), and the photo in the top left corner of IV contains white light. (C) Shows immunohistostaining results of BrdU (green) and CD31 (red) for cardiac tissues, and arrowhead indicates the location of LRCs after a 2 weeks chase. (D) Shows LRCs are mainly myocardium cells after a 3 months chase, and the asterisk in I–III shows BrdU (red) positive cells. (E) Shows LRCs located in the vicinity of vascular endothelial region with varied embedding after 3 months chase, and 6 asterisks indicate BrdU positive cells. (F) Shows identified LRCs in cardiac tissues after 24 months chase, and asterisk indicates BrdU positive myocardium cells, however, an arrow indicates ‘BrdU light’ myocardium cells. (G) Shows statistical analysis of BrdU label-retaining cells. The black boxes represent the mean values of LRCs (n = 2 mice at each time point). The error bars represent standard derivations. P = 0.026<0.05 shows there was significant difference (2 weeks versus 6 months), however, P = 0.103>0.05 indicates there was no significant difference (6 months versus 24 months). (H) Shows a model of LRCs in heart after birth, a part of activated-LRCs turn into d-LRCs. Scale bars: BI, BIII, C, E and F = 20 μm; DI–DIII = 30 μm; BII, BIV =  40 μm.
© Copyright Policy - open-access
Related In: Results  -  Collection

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

Figure 1: Existence and turnover of LRCs in cardiac tissues of normal adult mice(A) Shows schematic outlines of the BrdU pulse chase. w: week, m: month. (BI–IV) Show distinct locations of LRCs in the cardiac tissues after a 2 weeks chase, and the asterisk in I–IV indicates BrdU positive cells (asterisk), and the photo in the top left corner of IV contains white light. (C) Shows immunohistostaining results of BrdU (green) and CD31 (red) for cardiac tissues, and arrowhead indicates the location of LRCs after a 2 weeks chase. (D) Shows LRCs are mainly myocardium cells after a 3 months chase, and the asterisk in I–III shows BrdU (red) positive cells. (E) Shows LRCs located in the vicinity of vascular endothelial region with varied embedding after 3 months chase, and 6 asterisks indicate BrdU positive cells. (F) Shows identified LRCs in cardiac tissues after 24 months chase, and asterisk indicates BrdU positive myocardium cells, however, an arrow indicates ‘BrdU light’ myocardium cells. (G) Shows statistical analysis of BrdU label-retaining cells. The black boxes represent the mean values of LRCs (n = 2 mice at each time point). The error bars represent standard derivations. P = 0.026<0.05 shows there was significant difference (2 weeks versus 6 months), however, P = 0.103>0.05 indicates there was no significant difference (6 months versus 24 months). (H) Shows a model of LRCs in heart after birth, a part of activated-LRCs turn into d-LRCs. Scale bars: BI, BIII, C, E and F = 20 μm; DI–DIII = 30 μm; BII, BIV =  40 μm.
Mentions: Newborn mice (7 days old) were injected subcutaneously with BrdU (Sigma, pH 7.4) twice a day for 3–5 consecutive days (50–100 mg/kg per injection for each mouse). The labelled mice were killed at 2 weeks (n = 2 mice), 3 months (n = 2 mice), 6 months (n = 2 mice) and 24 months (n = 2 mice) (Figure 1A) to trace and locate the LRCs throughout their lifespan. The traced time was calculated from the initial administration of BrdU. According to Urbanek et al. (2006), cells with BrdU label can be divided into ‘BrdU bright’ cells and ‘BrdU dim’ cells.

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