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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

Chasing and kinetics of activated CSCs/CPCs every 12 h on day 4 post-injury(A) Shows schematic outlines of 5-Fu-induced injury plus BrdU pulse-chase assays. (B) Shows staining at the initiation of pulse chase, and the right corner photo in (B) shows vascular endothelial location of ‘BrdU bright’ cells; the dotted line indicates the VE region. (C) Shows staining after a 12 h chase. (D) Shows staining after a 24 h chase. (E) Shows staining after a 36 h chase. Nuclei were stained with DAPI (scale bar = 10 μm).
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Figure 3: Chasing and kinetics of activated CSCs/CPCs every 12 h on day 4 post-injury(A) Shows schematic outlines of 5-Fu-induced injury plus BrdU pulse-chase assays. (B) Shows staining at the initiation of pulse chase, and the right corner photo in (B) shows vascular endothelial location of ‘BrdU bright’ cells; the dotted line indicates the VE region. (C) Shows staining after a 12 h chase. (D) Shows staining after a 24 h chase. (E) Shows staining after a 36 h chase. Nuclei were stained with DAPI (scale bar = 10 μm).

Mentions: To chase the activated-CSCs/CPCs further in vivo, 5-Fu induced injury plus BrdU pulse was subsequently designed, and a chase of every 12 h was done through day 4 (Figure 3A). Immunohistostaining of BrdU and N-cad of cardiac sections was carried out on the material collected. At the initial BrdU pulse (0 h), most of the BrdU bright cells were located at the myocardium, including the endothelial region (Figure 3B), which suggested that the endothelial progenitor cells were located in the vascular endothelial region and they could take part in the regeneration of the injured heart. After a 12 h chase, most LRCs still appeared in the myocardium with scattered distribution (Figure 3C). Furthermore, the nuclear morphology of the BrdU bright cells at this time was similar to that in 0 h. However, after a 24 h chase, some LRCs with nuclear morphology of mature cardiomyocytes appeared in the myocardium (Figure 3D). After a 36 h chase, some LRCs together with surrounding cardiomyocytes had formed an embellished dish connection (Figure 3E).


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

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

Chasing and kinetics of activated CSCs/CPCs every 12 h on day 4 post-injury(A) Shows schematic outlines of 5-Fu-induced injury plus BrdU pulse-chase assays. (B) Shows staining at the initiation of pulse chase, and the right corner photo in (B) shows vascular endothelial location of ‘BrdU bright’ cells; the dotted line indicates the VE region. (C) Shows staining after a 12 h chase. (D) Shows staining after a 24 h chase. (E) Shows staining after a 36 h chase. Nuclei were stained with DAPI (scale bar = 10 μm).
© Copyright Policy - open-access
Related In: Results  -  Collection

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

Figure 3: Chasing and kinetics of activated CSCs/CPCs every 12 h on day 4 post-injury(A) Shows schematic outlines of 5-Fu-induced injury plus BrdU pulse-chase assays. (B) Shows staining at the initiation of pulse chase, and the right corner photo in (B) shows vascular endothelial location of ‘BrdU bright’ cells; the dotted line indicates the VE region. (C) Shows staining after a 12 h chase. (D) Shows staining after a 24 h chase. (E) Shows staining after a 36 h chase. Nuclei were stained with DAPI (scale bar = 10 μm).
Mentions: To chase the activated-CSCs/CPCs further in vivo, 5-Fu induced injury plus BrdU pulse was subsequently designed, and a chase of every 12 h was done through day 4 (Figure 3A). Immunohistostaining of BrdU and N-cad of cardiac sections was carried out on the material collected. At the initial BrdU pulse (0 h), most of the BrdU bright cells were located at the myocardium, including the endothelial region (Figure 3B), which suggested that the endothelial progenitor cells were located in the vascular endothelial region and they could take part in the regeneration of the injured heart. After a 12 h chase, most LRCs still appeared in the myocardium with scattered distribution (Figure 3C). Furthermore, the nuclear morphology of the BrdU bright cells at this time was similar to that in 0 h. However, after a 24 h chase, some LRCs with nuclear morphology of mature cardiomyocytes appeared in the myocardium (Figure 3D). After a 36 h chase, some LRCs together with surrounding cardiomyocytes had formed an embellished dish connection (Figure 3E).

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