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Xenotransplantation of human adipose-derived stem cells in zebrafish embryos.

Li J, Zeng G, Qi Y, Tang X, Zhang J, Wu Z, Liang J, Shi L, Liu H, Zhang P - PLoS ONE (2015)

Bottom Line: The results indicated that human ADSCs did not change their cell viability and the expression levels of cell surface antigens after GFP transduction.The expression of CD105 was observable in the xenotransplanted ADSCs, but CD31 expression was undetectable.Therefore, our results indicate that human ADSCs xenotransplanted in the zebrafish embryos not only can survive and proliferate at across-species circumstance, but also seem to maintain their undifferentiation status in a short term.

View Article: PubMed Central - PubMed

Affiliation: Institute of Plastic Surgery, Affiliated Hospital of Guangdong Medical College, Zhanjiang, Guangdong Province, China.

ABSTRACT
Zebrafish is a widely used animal model with well-characterized background in developmental biology. The fate of human adipose-derived stem cells (ADSCs) after their xenotransplantation into the developing embryos of zebrafish is unknown. Therefore, human ADSCs were firstly isolated, and then transduced with lentiviral vector system carrying a green fluorescent protein (GFP) reporter gene, and followed by detection of their cell viability and the expression of cell surface antigens. These GFP-expressing human ADSCs were transplanted into the zebrafish embryos at 3.3-4.3 hour post-fertilization (hpf). Green fluorescent signal, the proliferation and differentiation of human ADSCs in recipient embryos were respectively examined using fluorescent microscopy and immunohistochemical staining. The results indicated that human ADSCs did not change their cell viability and the expression levels of cell surface antigens after GFP transduction. Microscopic examination demonstrated that green fluorescent signals of GFP expressed in the transplanted cells were observed in the embryos and larva fish at post-transplantation. The positive staining of Ki-67 revealed the survival and proliferation of human ADSCs in fish larvae after transplantation. The expression of CD105 was observable in the xenotransplanted ADSCs, but CD31 expression was undetectable. Therefore, our results indicate that human ADSCs xenotransplanted in the zebrafish embryos not only can survive and proliferate at across-species circumstance, but also seem to maintain their undifferentiation status in a short term. This xenograft model of zebrafish embryos may provide a promising and useful technical platform for the investigation of biology and physiology of stem cells in vivo.

No MeSH data available.


Related in: MedlinePlus

Detection of cell surface antigens.Cell surface antigens of GFP-expressing ADSCs and control ADSCs were detected with flow cytometry. Nonspecific IgG was served as the control of background fluorescence. The positive percentage of cell surface was showed in the histograms.
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pone.0123264.g003: Detection of cell surface antigens.Cell surface antigens of GFP-expressing ADSCs and control ADSCs were detected with flow cytometry. Nonspecific IgG was served as the control of background fluorescence. The positive percentage of cell surface was showed in the histograms.

Mentions: As shown in Fig 3, high levels of CD marker CD29 (98.20%), CD44 (97.30%), CD90 (96.60%), and CD105 (89.80%) in GFP-expressing ADSCs after transduction were displayed, and they were considered as the markers for mesenchymal stem cells [30]. Similarly, these high levels of CD markers also could be detected in the control ADSCs. In contrast, the positives percentage of hematopoietic lineage markers CD31, CD34, and CD45 were under 2.0% in the GFP-expressing ADSCs or hardly detectable in the control ADSCs. Additionally, the results of flow cytometry analysis also showed a high level of CD13 expression and the absence of CD14 and CD106 expression in the GFP-expressing ADSCs and control cells, consistent with previous reports [31].


Xenotransplantation of human adipose-derived stem cells in zebrafish embryos.

Li J, Zeng G, Qi Y, Tang X, Zhang J, Wu Z, Liang J, Shi L, Liu H, Zhang P - PLoS ONE (2015)

Detection of cell surface antigens.Cell surface antigens of GFP-expressing ADSCs and control ADSCs were detected with flow cytometry. Nonspecific IgG was served as the control of background fluorescence. The positive percentage of cell surface was showed in the histograms.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4388839&req=5

pone.0123264.g003: Detection of cell surface antigens.Cell surface antigens of GFP-expressing ADSCs and control ADSCs were detected with flow cytometry. Nonspecific IgG was served as the control of background fluorescence. The positive percentage of cell surface was showed in the histograms.
Mentions: As shown in Fig 3, high levels of CD marker CD29 (98.20%), CD44 (97.30%), CD90 (96.60%), and CD105 (89.80%) in GFP-expressing ADSCs after transduction were displayed, and they were considered as the markers for mesenchymal stem cells [30]. Similarly, these high levels of CD markers also could be detected in the control ADSCs. In contrast, the positives percentage of hematopoietic lineage markers CD31, CD34, and CD45 were under 2.0% in the GFP-expressing ADSCs or hardly detectable in the control ADSCs. Additionally, the results of flow cytometry analysis also showed a high level of CD13 expression and the absence of CD14 and CD106 expression in the GFP-expressing ADSCs and control cells, consistent with previous reports [31].

Bottom Line: The results indicated that human ADSCs did not change their cell viability and the expression levels of cell surface antigens after GFP transduction.The expression of CD105 was observable in the xenotransplanted ADSCs, but CD31 expression was undetectable.Therefore, our results indicate that human ADSCs xenotransplanted in the zebrafish embryos not only can survive and proliferate at across-species circumstance, but also seem to maintain their undifferentiation status in a short term.

View Article: PubMed Central - PubMed

Affiliation: Institute of Plastic Surgery, Affiliated Hospital of Guangdong Medical College, Zhanjiang, Guangdong Province, China.

ABSTRACT
Zebrafish is a widely used animal model with well-characterized background in developmental biology. The fate of human adipose-derived stem cells (ADSCs) after their xenotransplantation into the developing embryos of zebrafish is unknown. Therefore, human ADSCs were firstly isolated, and then transduced with lentiviral vector system carrying a green fluorescent protein (GFP) reporter gene, and followed by detection of their cell viability and the expression of cell surface antigens. These GFP-expressing human ADSCs were transplanted into the zebrafish embryos at 3.3-4.3 hour post-fertilization (hpf). Green fluorescent signal, the proliferation and differentiation of human ADSCs in recipient embryos were respectively examined using fluorescent microscopy and immunohistochemical staining. The results indicated that human ADSCs did not change their cell viability and the expression levels of cell surface antigens after GFP transduction. Microscopic examination demonstrated that green fluorescent signals of GFP expressed in the transplanted cells were observed in the embryos and larva fish at post-transplantation. The positive staining of Ki-67 revealed the survival and proliferation of human ADSCs in fish larvae after transplantation. The expression of CD105 was observable in the xenotransplanted ADSCs, but CD31 expression was undetectable. Therefore, our results indicate that human ADSCs xenotransplanted in the zebrafish embryos not only can survive and proliferate at across-species circumstance, but also seem to maintain their undifferentiation status in a short term. This xenograft model of zebrafish embryos may provide a promising and useful technical platform for the investigation of biology and physiology of stem cells in vivo.

No MeSH data available.


Related in: MedlinePlus