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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 CD105 and CD31 expression of ADSCs.Human ADSCs were xenotransplanted into the zebrafish embryos at 3.3–4.3hpf, and immunohistochemical staining and immunocytochemical staining were used to evaluate the expression of CD105 and CD31 of human ADSCs in the zebrafish at 48 hpf, and in culture dishes. (A-D) Representative images of immunohistochemical staining: the xenotransplanted ADSCs expressed positive CD105(brownish, indicated by green arrow) in the zebrafish embryos (A), and CD105 expression could not be detected in the control without ADSCs transplantation (B); the xenotransplanted ADSCs (indicated by red arrow) did not express CD31 antigen in the zebrafish embryos (C), and CD31could not be detected in the control without ADSCs transplantation (D). (E-H) Representative images of immunocytochemical staining: CD105 expression was detectable in the ADSCs cultured in the dishes before transplantation (E), and negative control for CD105 (without the addition of primary antibody) (F); CD31 expression was not detectable in the ADSCs cultured in the dishes before transplantation (G) and in the negative control (H). Scale bar: 100 μm. ab(+): added primary antibody; ab(-): no primary antibody.
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pone.0123264.g008: Detection of CD105 and CD31 expression of ADSCs.Human ADSCs were xenotransplanted into the zebrafish embryos at 3.3–4.3hpf, and immunohistochemical staining and immunocytochemical staining were used to evaluate the expression of CD105 and CD31 of human ADSCs in the zebrafish at 48 hpf, and in culture dishes. (A-D) Representative images of immunohistochemical staining: the xenotransplanted ADSCs expressed positive CD105(brownish, indicated by green arrow) in the zebrafish embryos (A), and CD105 expression could not be detected in the control without ADSCs transplantation (B); the xenotransplanted ADSCs (indicated by red arrow) did not express CD31 antigen in the zebrafish embryos (C), and CD31could not be detected in the control without ADSCs transplantation (D). (E-H) Representative images of immunocytochemical staining: CD105 expression was detectable in the ADSCs cultured in the dishes before transplantation (E), and negative control for CD105 (without the addition of primary antibody) (F); CD31 expression was not detectable in the ADSCs cultured in the dishes before transplantation (G) and in the negative control (H). Scale bar: 100 μm. ab(+): added primary antibody; ab(-): no primary antibody.

Mentions: Immunohistochemical staining analysis indicated that cell surface antigen CD105 expression was observed in the xenotransplanted human ADSCs in zebrafish embryos at 48 hpf, and the human ADSCs cultured in the dishes also expressed CD105. However, CD31expression could not be detected in the recipient zebrafish and cultured ADSCs in the dishes (Fig 8). The immunofluorescence staining for the detection of CD105 and CD31 expression further confirmed that the transplanted ADSCs in the zebrafish could express CD105, but not CD31 (Fig 9). These results seem to suggest that human ADSCs can keep their property and maintain the state of undifferentiation in a short term.


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 CD105 and CD31 expression of ADSCs.Human ADSCs were xenotransplanted into the zebrafish embryos at 3.3–4.3hpf, and immunohistochemical staining and immunocytochemical staining were used to evaluate the expression of CD105 and CD31 of human ADSCs in the zebrafish at 48 hpf, and in culture dishes. (A-D) Representative images of immunohistochemical staining: the xenotransplanted ADSCs expressed positive CD105(brownish, indicated by green arrow) in the zebrafish embryos (A), and CD105 expression could not be detected in the control without ADSCs transplantation (B); the xenotransplanted ADSCs (indicated by red arrow) did not express CD31 antigen in the zebrafish embryos (C), and CD31could not be detected in the control without ADSCs transplantation (D). (E-H) Representative images of immunocytochemical staining: CD105 expression was detectable in the ADSCs cultured in the dishes before transplantation (E), and negative control for CD105 (without the addition of primary antibody) (F); CD31 expression was not detectable in the ADSCs cultured in the dishes before transplantation (G) and in the negative control (H). Scale bar: 100 μm. ab(+): added primary antibody; ab(-): no primary antibody.
© Copyright Policy
Related In: Results  -  Collection

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

pone.0123264.g008: Detection of CD105 and CD31 expression of ADSCs.Human ADSCs were xenotransplanted into the zebrafish embryos at 3.3–4.3hpf, and immunohistochemical staining and immunocytochemical staining were used to evaluate the expression of CD105 and CD31 of human ADSCs in the zebrafish at 48 hpf, and in culture dishes. (A-D) Representative images of immunohistochemical staining: the xenotransplanted ADSCs expressed positive CD105(brownish, indicated by green arrow) in the zebrafish embryos (A), and CD105 expression could not be detected in the control without ADSCs transplantation (B); the xenotransplanted ADSCs (indicated by red arrow) did not express CD31 antigen in the zebrafish embryos (C), and CD31could not be detected in the control without ADSCs transplantation (D). (E-H) Representative images of immunocytochemical staining: CD105 expression was detectable in the ADSCs cultured in the dishes before transplantation (E), and negative control for CD105 (without the addition of primary antibody) (F); CD31 expression was not detectable in the ADSCs cultured in the dishes before transplantation (G) and in the negative control (H). Scale bar: 100 μm. ab(+): added primary antibody; ab(-): no primary antibody.
Mentions: Immunohistochemical staining analysis indicated that cell surface antigen CD105 expression was observed in the xenotransplanted human ADSCs in zebrafish embryos at 48 hpf, and the human ADSCs cultured in the dishes also expressed CD105. However, CD31expression could not be detected in the recipient zebrafish and cultured ADSCs in the dishes (Fig 8). The immunofluorescence staining for the detection of CD105 and CD31 expression further confirmed that the transplanted ADSCs in the zebrafish could express CD105, but not CD31 (Fig 9). These results seem to suggest that human ADSCs can keep their property and maintain the state of undifferentiation in a short term.

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