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Identification and isolation of kidney-derived stem cells from transgenic rats with diphtheria toxin-induced kidney damage

View Article: PubMed Central - PubMed

ABSTRACT

Adult stem cells have been well characterized in numerous organs, with the exception of the kidneys. Therefore, the present study aimed to identify and isolate kidney-derived stem cells. A total of 12 Fischer 344 transgenic rats expressing the human diphtheria toxin receptor in podocyte cells of the kidney, were used in the present study. The rats were administered 5-bromo-2′-deoxyuridine (BrdU) in order to detect cellular proliferation. After 60 days, the rats were treated with the diphtheria toxin (DT), in order to induce kidney injury. Immunohistochemical analysis indicated that the number of BrdU-positive cells were increased following DT treatment. In addition, the expression of octamer-binding transcription factor 4 (Oct-4), a stem cell marker, was detected and suggested that kidney-specific stem cells were present in the DT-treated tissue samples. Furthermore, tissue samples exhibited repair of the DT-induced injury. Further cellular culturing was conducted in order to isolate the kidney-specific stem cells. After 5 weeks of culture, the majority of the cells were non-viable, with the exception of certain specialized, unique cell types, which were monomorphic and spindle-shaped in appearance. The unique cells were isolated and subjected to immunostaining and reverse transcription-polymerase chain reaction analyses in order to reconfirm the expression of Oct-4 and to detect the expression of Paired box 2 (Pax-2), which is necessary for the formation of kidney structures. The unique cells were positive for Oct-4 and Pax-2; thus suggesting that the identified cells were kidney-derived stem cells. The results of the present study suggested that the unique cell type identified in the kidneys of the DT-treated rats were kidney-specific stem cells that may have been involved in the repair of DT-induced tissue injury. In addition, these cells may provide a useful cell line for studying the fundamental characteristics of kidney stem cells, as well as identifying kidney-specific stem cell markers.

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5-bromo-2′-deoxyuridine (BrdU) labeling retention assay. (A) BrdU-treated control Tg rat kidney tissue section. The BrdU-positive cells (red) are marked with white arrows. (B) BrdU + diphtheria toxin (DT)-treated Tg rat kidney tissue section. DT-treated cells are marked with yellow arrows (scale bar, 50 µm).
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f1-etm-0-0-3516: 5-bromo-2′-deoxyuridine (BrdU) labeling retention assay. (A) BrdU-treated control Tg rat kidney tissue section. The BrdU-positive cells (red) are marked with white arrows. (B) BrdU + diphtheria toxin (DT)-treated Tg rat kidney tissue section. DT-treated cells are marked with yellow arrows (scale bar, 50 µm).

Mentions: Kidney samples were collected from the Tg rats (including the control rats) and immunohistochemical analyses were conducted using anti-BrdU and anti-Oct-4 antibodies. BrdU-positive cells were detected in the control and DT-treated kidney tissue sections; however, the number of BrdU-positive cells in the control sections was markedly decreased, as compared with the DT-treated kidney sections (Fig. 1A and B). In addition, the DT-treated cells appeared to be renewed by the BrdU-positive cells (Fig. 1B), and these results were confirmed by immunohistochemical detection of Oct-4 and BrdU co-expression. Therefore, these results suggested that the majority of the BrdU-positive cells were kidney-derived stem cells, since they also expressed Oct-4 (Fig. 2). These kidney-derived stem cells may be involved in the restoration or renewal of the injured or depleted cells.


Identification and isolation of kidney-derived stem cells from transgenic rats with diphtheria toxin-induced kidney damage
5-bromo-2′-deoxyuridine (BrdU) labeling retention assay. (A) BrdU-treated control Tg rat kidney tissue section. The BrdU-positive cells (red) are marked with white arrows. (B) BrdU + diphtheria toxin (DT)-treated Tg rat kidney tissue section. DT-treated cells are marked with yellow arrows (scale bar, 50 µm).
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4998113&req=5

f1-etm-0-0-3516: 5-bromo-2′-deoxyuridine (BrdU) labeling retention assay. (A) BrdU-treated control Tg rat kidney tissue section. The BrdU-positive cells (red) are marked with white arrows. (B) BrdU + diphtheria toxin (DT)-treated Tg rat kidney tissue section. DT-treated cells are marked with yellow arrows (scale bar, 50 µm).
Mentions: Kidney samples were collected from the Tg rats (including the control rats) and immunohistochemical analyses were conducted using anti-BrdU and anti-Oct-4 antibodies. BrdU-positive cells were detected in the control and DT-treated kidney tissue sections; however, the number of BrdU-positive cells in the control sections was markedly decreased, as compared with the DT-treated kidney sections (Fig. 1A and B). In addition, the DT-treated cells appeared to be renewed by the BrdU-positive cells (Fig. 1B), and these results were confirmed by immunohistochemical detection of Oct-4 and BrdU co-expression. Therefore, these results suggested that the majority of the BrdU-positive cells were kidney-derived stem cells, since they also expressed Oct-4 (Fig. 2). These kidney-derived stem cells may be involved in the restoration or renewal of the injured or depleted cells.

View Article: PubMed Central - PubMed

ABSTRACT

Adult stem cells have been well characterized in numerous organs, with the exception of the kidneys. Therefore, the present study aimed to identify and isolate kidney-derived stem cells. A total of 12 Fischer 344 transgenic rats expressing the human diphtheria toxin receptor in podocyte cells of the kidney, were used in the present study. The rats were administered 5-bromo-2′-deoxyuridine (BrdU) in order to detect cellular proliferation. After 60 days, the rats were treated with the diphtheria toxin (DT), in order to induce kidney injury. Immunohistochemical analysis indicated that the number of BrdU-positive cells were increased following DT treatment. In addition, the expression of octamer-binding transcription factor 4 (Oct-4), a stem cell marker, was detected and suggested that kidney-specific stem cells were present in the DT-treated tissue samples. Furthermore, tissue samples exhibited repair of the DT-induced injury. Further cellular culturing was conducted in order to isolate the kidney-specific stem cells. After 5 weeks of culture, the majority of the cells were non-viable, with the exception of certain specialized, unique cell types, which were monomorphic and spindle-shaped in appearance. The unique cells were isolated and subjected to immunostaining and reverse transcription-polymerase chain reaction analyses in order to reconfirm the expression of Oct-4 and to detect the expression of Paired box 2 (Pax-2), which is necessary for the formation of kidney structures. The unique cells were positive for Oct-4 and Pax-2; thus suggesting that the identified cells were kidney-derived stem cells. The results of the present study suggested that the unique cell type identified in the kidneys of the DT-treated rats were kidney-specific stem cells that may have been involved in the repair of DT-induced tissue injury. In addition, these cells may provide a useful cell line for studying the fundamental characteristics of kidney stem cells, as well as identifying kidney-specific stem cell markers.

No MeSH data available.


Related in: MedlinePlus