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Amniotic fluid stem cells provide considerable advantages in epidermal regeneration: B7H4 creates a moderate inflammation microenvironment to promote wound repair.

Sun Q, Li F, Li H, Chen RH, Gu YZ, Chen Y, Liang HS, You XR, Ding SS, Gao L, Wang YL, Qin MD, Zhang XG - Sci Rep (2015)

Bottom Line: Co-localization of GFP/K14 and GFP/K10 in the repaired epidermis demonstrated that hAFS cells can differentiate into keratinocytes.During wound repair, hAFS cells did not directly secrete repair-related factors, such as bFGF, VEGF, CXCL12, TGF-β1 and KGF, and provided a moderate inflammation reaction with lower expression of IL-1β, IL-6, TNF-α, Cox2 and Mac3.Furthermore, with their uniquely high proliferation rate, hAFS cells offer a promising alternative for epidermal regeneration.

View Article: PubMed Central - PubMed

Affiliation: 1] The Stem Cell and Biomedical Material Key Laboratory of Jiangsu Province (the State Key Laboratory Incubation Base), Soochow University, Suzhou, Jiangsu Province, P.R. China [2] Department of Immunology, School of Biology and Basic Medical Sciences, Soochow University, Suzhou, Jiangsu Province, P.R. China.

ABSTRACT
The current treatments for severe skin injury all involve skin grafting. However, there is a worldwide shortage of donor skin tissue. In this study, we examined the advantages of using human amniotic fluid stem (hAFS) cells in skin wound healing. In vitro, hAFS cells differentiate into keratinocytes (termed hAFS-K). Like keratinocytes, hAFS-K cells express the markers K5, K14, K10 and involucrin; display typical cellular structure, including a tonofibril-rich cytoplasm; and construct a completely pluristratified epithelium in 3D culture. In vivo, in a mouse excisional wound model, GFP-positive hAFS cells participate in wound repair. Co-localization of GFP/K14 and GFP/K10 in the repaired epidermis demonstrated that hAFS cells can differentiate into keratinocytes. Real-time PCR results confirmed that hAFS cells can initiate and promote early-stage repair of skin damage. During wound repair, hAFS cells did not directly secrete repair-related factors, such as bFGF, VEGF, CXCL12, TGF-β1 and KGF, and provided a moderate inflammation reaction with lower expression of IL-1β, IL-6, TNF-α, Cox2 and Mac3. In hAFS cells, the negative co-stimulatory molecule B7H4 regulates low immunogenicity, which can provide a modest inflammatory reaction microenvironment for wound repair. Furthermore, with their uniquely high proliferation rate, hAFS cells offer a promising alternative for epidermal regeneration.

No MeSH data available.


Related in: MedlinePlus

GFP-positive hAFS cells directly promote and contribute to wound healing in vivo(a) Representative photographs of the wounds with GFP-positive hAFS cells, fibroblasts or sham cells at day 0, day 7, day 14 and day 21 are shown. (b) Measurement of wounds remaining in the sham, fibroblast, and hAFS cell groups in BALB/c mice. Analysis of variance (ANOVA) versus sham or fibroblast groups. The wounds were measured using the UTHSCSA ImageTool; *p < 0.01. (c) Histological analysis of wounds on BALB/c mice in the sham, fibroblast and hAFS groups at days 4 and 14 (H&E stain). At day 4, the wound edge is indicated by the black dashed lines. The infiltrated inflammatory cells (black arrowheads) and the capillary-like structures (red arrowheads) are indicated in the granulation tissue. At day 14, the repaired skin area is indicated by the black dashed lines, and newly formed hair follicles (blue arrowheads) are also shown. Abbreviations: M, muscle; F, hair follicle; P, papillae. (d) Flow cytometry of GFP-positive cells in the wound at 7, 14 and 21 days after wounding. (e) Immunofluorescence analysis of the presence of GFP-positive hAFS cells and fibroblasts in the wound at 7, 14 and 21 days. (f) Tissue sections from hAFS cell-treated wounds at day 7 were stained with K14 and K10 antibodies in red. The co-localization of K14 with GFP and K10 with GFP in the merged images appears orange.
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f4: GFP-positive hAFS cells directly promote and contribute to wound healing in vivo(a) Representative photographs of the wounds with GFP-positive hAFS cells, fibroblasts or sham cells at day 0, day 7, day 14 and day 21 are shown. (b) Measurement of wounds remaining in the sham, fibroblast, and hAFS cell groups in BALB/c mice. Analysis of variance (ANOVA) versus sham or fibroblast groups. The wounds were measured using the UTHSCSA ImageTool; *p < 0.01. (c) Histological analysis of wounds on BALB/c mice in the sham, fibroblast and hAFS groups at days 4 and 14 (H&E stain). At day 4, the wound edge is indicated by the black dashed lines. The infiltrated inflammatory cells (black arrowheads) and the capillary-like structures (red arrowheads) are indicated in the granulation tissue. At day 14, the repaired skin area is indicated by the black dashed lines, and newly formed hair follicles (blue arrowheads) are also shown. Abbreviations: M, muscle; F, hair follicle; P, papillae. (d) Flow cytometry of GFP-positive cells in the wound at 7, 14 and 21 days after wounding. (e) Immunofluorescence analysis of the presence of GFP-positive hAFS cells and fibroblasts in the wound at 7, 14 and 21 days. (f) Tissue sections from hAFS cell-treated wounds at day 7 were stained with K14 and K10 antibodies in red. The co-localization of K14 with GFP and K10 with GFP in the merged images appears orange.

Mentions: A GFP reporter gene was introduced into hAFS cells and fibroblasts using a lentiviral vector to enable the cells to be traced in the subsequent experiments. Flow cytometry and immunofluorescence showed that almost 100% of hAFS cells and fibroblasts were GFP positive (Suppl. Fig. S1a). We used an excision wound model in BALB/c mice with a ring to reduce self-repair, as previously described27. GFP-positive hAFS cells were injected into the wound bed. Over time, wounds treated with hAFS cells exhibited accelerated wound closure when compared to fibroblast-treated wounds or sham groups (Fig. 4a and Suppl. Fig. S1b). At day 7, there was more wound healing in hAFS cell-treated mice than the fibroblast and sham groups. At day 21, the wounds in hAFS cell-treated mice (n = 12) achieved almost complete wound closure, whereas no completely closed wounds were observed in the fibroblast-treated (n = 8) or sham group (n = 7) mice. These results show that hAFS cells can quickly and efficiently promote wound healing (Fig. 4b).


Amniotic fluid stem cells provide considerable advantages in epidermal regeneration: B7H4 creates a moderate inflammation microenvironment to promote wound repair.

Sun Q, Li F, Li H, Chen RH, Gu YZ, Chen Y, Liang HS, You XR, Ding SS, Gao L, Wang YL, Qin MD, Zhang XG - Sci Rep (2015)

GFP-positive hAFS cells directly promote and contribute to wound healing in vivo(a) Representative photographs of the wounds with GFP-positive hAFS cells, fibroblasts or sham cells at day 0, day 7, day 14 and day 21 are shown. (b) Measurement of wounds remaining in the sham, fibroblast, and hAFS cell groups in BALB/c mice. Analysis of variance (ANOVA) versus sham or fibroblast groups. The wounds were measured using the UTHSCSA ImageTool; *p < 0.01. (c) Histological analysis of wounds on BALB/c mice in the sham, fibroblast and hAFS groups at days 4 and 14 (H&E stain). At day 4, the wound edge is indicated by the black dashed lines. The infiltrated inflammatory cells (black arrowheads) and the capillary-like structures (red arrowheads) are indicated in the granulation tissue. At day 14, the repaired skin area is indicated by the black dashed lines, and newly formed hair follicles (blue arrowheads) are also shown. Abbreviations: M, muscle; F, hair follicle; P, papillae. (d) Flow cytometry of GFP-positive cells in the wound at 7, 14 and 21 days after wounding. (e) Immunofluorescence analysis of the presence of GFP-positive hAFS cells and fibroblasts in the wound at 7, 14 and 21 days. (f) Tissue sections from hAFS cell-treated wounds at day 7 were stained with K14 and K10 antibodies in red. The co-localization of K14 with GFP and K10 with GFP in the merged images appears orange.
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Related In: Results  -  Collection

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f4: GFP-positive hAFS cells directly promote and contribute to wound healing in vivo(a) Representative photographs of the wounds with GFP-positive hAFS cells, fibroblasts or sham cells at day 0, day 7, day 14 and day 21 are shown. (b) Measurement of wounds remaining in the sham, fibroblast, and hAFS cell groups in BALB/c mice. Analysis of variance (ANOVA) versus sham or fibroblast groups. The wounds were measured using the UTHSCSA ImageTool; *p < 0.01. (c) Histological analysis of wounds on BALB/c mice in the sham, fibroblast and hAFS groups at days 4 and 14 (H&E stain). At day 4, the wound edge is indicated by the black dashed lines. The infiltrated inflammatory cells (black arrowheads) and the capillary-like structures (red arrowheads) are indicated in the granulation tissue. At day 14, the repaired skin area is indicated by the black dashed lines, and newly formed hair follicles (blue arrowheads) are also shown. Abbreviations: M, muscle; F, hair follicle; P, papillae. (d) Flow cytometry of GFP-positive cells in the wound at 7, 14 and 21 days after wounding. (e) Immunofluorescence analysis of the presence of GFP-positive hAFS cells and fibroblasts in the wound at 7, 14 and 21 days. (f) Tissue sections from hAFS cell-treated wounds at day 7 were stained with K14 and K10 antibodies in red. The co-localization of K14 with GFP and K10 with GFP in the merged images appears orange.
Mentions: A GFP reporter gene was introduced into hAFS cells and fibroblasts using a lentiviral vector to enable the cells to be traced in the subsequent experiments. Flow cytometry and immunofluorescence showed that almost 100% of hAFS cells and fibroblasts were GFP positive (Suppl. Fig. S1a). We used an excision wound model in BALB/c mice with a ring to reduce self-repair, as previously described27. GFP-positive hAFS cells were injected into the wound bed. Over time, wounds treated with hAFS cells exhibited accelerated wound closure when compared to fibroblast-treated wounds or sham groups (Fig. 4a and Suppl. Fig. S1b). At day 7, there was more wound healing in hAFS cell-treated mice than the fibroblast and sham groups. At day 21, the wounds in hAFS cell-treated mice (n = 12) achieved almost complete wound closure, whereas no completely closed wounds were observed in the fibroblast-treated (n = 8) or sham group (n = 7) mice. These results show that hAFS cells can quickly and efficiently promote wound healing (Fig. 4b).

Bottom Line: Co-localization of GFP/K14 and GFP/K10 in the repaired epidermis demonstrated that hAFS cells can differentiate into keratinocytes.During wound repair, hAFS cells did not directly secrete repair-related factors, such as bFGF, VEGF, CXCL12, TGF-β1 and KGF, and provided a moderate inflammation reaction with lower expression of IL-1β, IL-6, TNF-α, Cox2 and Mac3.Furthermore, with their uniquely high proliferation rate, hAFS cells offer a promising alternative for epidermal regeneration.

View Article: PubMed Central - PubMed

Affiliation: 1] The Stem Cell and Biomedical Material Key Laboratory of Jiangsu Province (the State Key Laboratory Incubation Base), Soochow University, Suzhou, Jiangsu Province, P.R. China [2] Department of Immunology, School of Biology and Basic Medical Sciences, Soochow University, Suzhou, Jiangsu Province, P.R. China.

ABSTRACT
The current treatments for severe skin injury all involve skin grafting. However, there is a worldwide shortage of donor skin tissue. In this study, we examined the advantages of using human amniotic fluid stem (hAFS) cells in skin wound healing. In vitro, hAFS cells differentiate into keratinocytes (termed hAFS-K). Like keratinocytes, hAFS-K cells express the markers K5, K14, K10 and involucrin; display typical cellular structure, including a tonofibril-rich cytoplasm; and construct a completely pluristratified epithelium in 3D culture. In vivo, in a mouse excisional wound model, GFP-positive hAFS cells participate in wound repair. Co-localization of GFP/K14 and GFP/K10 in the repaired epidermis demonstrated that hAFS cells can differentiate into keratinocytes. Real-time PCR results confirmed that hAFS cells can initiate and promote early-stage repair of skin damage. During wound repair, hAFS cells did not directly secrete repair-related factors, such as bFGF, VEGF, CXCL12, TGF-β1 and KGF, and provided a moderate inflammation reaction with lower expression of IL-1β, IL-6, TNF-α, Cox2 and Mac3. In hAFS cells, the negative co-stimulatory molecule B7H4 regulates low immunogenicity, which can provide a modest inflammatory reaction microenvironment for wound repair. Furthermore, with their uniquely high proliferation rate, hAFS cells offer a promising alternative for epidermal regeneration.

No MeSH data available.


Related in: MedlinePlus