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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

The transplanted ADSCs express CD105, but not CD31 in zebrafish.The GFP-expressing human ADSCs were xenotransplanted into the zebrafish embryos at 3.3–4.3hpf, and immunofluorescence staining was performed to detect CD105 and CD31 expression of human ADSCs in the zebrafish at 48 hpf, and the images were captured with laser confocal scanning microscope. A-H: Representative images of immunofluorescence staining for zebrafish tissue sections. (A-D) Co-localization of CD105 and GFP in the transplanted ADSCs: (A) Bright field; (B) CD105 was positive; (C) GFP was positive; (D) Merged (A), (B) and (C). (E-H) The co-localization of CD31 and GFP in the transplanted ADSCs: (E) Bright field; (F) CD31 was negative; (G) GFP was positive; (H) Merged (E), (F) and (G). (I-N) Representative images of immunofluorescence staining for the ADSCs seeded on the glass slides. (I-K) The co-localization of CD105 and GFP in the human ADSCs before their transplantation: (I) CD105 expression was positive; (J) GFP was positive; (K) Merged (I) and (J). (L-N) The co-localization of CD31 and GFP in the human ADSCs before their transplantation: (L) CD31expression was negative; (M) GFP was positive; (N) Merged (L) and (M).
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pone.0123264.g009: The transplanted ADSCs express CD105, but not CD31 in zebrafish.The GFP-expressing human ADSCs were xenotransplanted into the zebrafish embryos at 3.3–4.3hpf, and immunofluorescence staining was performed to detect CD105 and CD31 expression of human ADSCs in the zebrafish at 48 hpf, and the images were captured with laser confocal scanning microscope. A-H: Representative images of immunofluorescence staining for zebrafish tissue sections. (A-D) Co-localization of CD105 and GFP in the transplanted ADSCs: (A) Bright field; (B) CD105 was positive; (C) GFP was positive; (D) Merged (A), (B) and (C). (E-H) The co-localization of CD31 and GFP in the transplanted ADSCs: (E) Bright field; (F) CD31 was negative; (G) GFP was positive; (H) Merged (E), (F) and (G). (I-N) Representative images of immunofluorescence staining for the ADSCs seeded on the glass slides. (I-K) The co-localization of CD105 and GFP in the human ADSCs before their transplantation: (I) CD105 expression was positive; (J) GFP was positive; (K) Merged (I) and (J). (L-N) The co-localization of CD31 and GFP in the human ADSCs before their transplantation: (L) CD31expression was negative; (M) GFP was positive; (N) Merged (L) and (M).

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)

The transplanted ADSCs express CD105, but not CD31 in zebrafish.The GFP-expressing human ADSCs were xenotransplanted into the zebrafish embryos at 3.3–4.3hpf, and immunofluorescence staining was performed to detect CD105 and CD31 expression of human ADSCs in the zebrafish at 48 hpf, and the images were captured with laser confocal scanning microscope. A-H: Representative images of immunofluorescence staining for zebrafish tissue sections. (A-D) Co-localization of CD105 and GFP in the transplanted ADSCs: (A) Bright field; (B) CD105 was positive; (C) GFP was positive; (D) Merged (A), (B) and (C). (E-H) The co-localization of CD31 and GFP in the transplanted ADSCs: (E) Bright field; (F) CD31 was negative; (G) GFP was positive; (H) Merged (E), (F) and (G). (I-N) Representative images of immunofluorescence staining for the ADSCs seeded on the glass slides. (I-K) The co-localization of CD105 and GFP in the human ADSCs before their transplantation: (I) CD105 expression was positive; (J) GFP was positive; (K) Merged (I) and (J). (L-N) The co-localization of CD31 and GFP in the human ADSCs before their transplantation: (L) CD31expression was negative; (M) GFP was positive; (N) Merged (L) and (M).
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pone.0123264.g009: The transplanted ADSCs express CD105, but not CD31 in zebrafish.The GFP-expressing human ADSCs were xenotransplanted into the zebrafish embryos at 3.3–4.3hpf, and immunofluorescence staining was performed to detect CD105 and CD31 expression of human ADSCs in the zebrafish at 48 hpf, and the images were captured with laser confocal scanning microscope. A-H: Representative images of immunofluorescence staining for zebrafish tissue sections. (A-D) Co-localization of CD105 and GFP in the transplanted ADSCs: (A) Bright field; (B) CD105 was positive; (C) GFP was positive; (D) Merged (A), (B) and (C). (E-H) The co-localization of CD31 and GFP in the transplanted ADSCs: (E) Bright field; (F) CD31 was negative; (G) GFP was positive; (H) Merged (E), (F) and (G). (I-N) Representative images of immunofluorescence staining for the ADSCs seeded on the glass slides. (I-K) The co-localization of CD105 and GFP in the human ADSCs before their transplantation: (I) CD105 expression was positive; (J) GFP was positive; (K) Merged (I) and (J). (L-N) The co-localization of CD31 and GFP in the human ADSCs before their transplantation: (L) CD31expression was negative; (M) GFP was positive; (N) Merged (L) and (M).
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