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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

Ultrastructural characteristics of hAFS cells and differentiation in 3D culture(a) Transmission electron microscope images of hAFS cells, hAFS-K and mature keratinocytes. Insets show tonofibrils in hAFS-K and keratinocytes (blue arrows), which are missing in hAFS cells. (b) H&E images of normal human skin and the epidermis constructed by hAFS-K demonstrate a well-organized, multilayer epithelium that contains all morphological layers (blue lines) and a collagen matrix in which resident fibroblasts (red line) play the role of the dermis. An epidermis constructed by keratinocytes was used as a positive control. (c) Immunohistochemistry of an organotypic epidermis constructed by hAFS-K showing a normal distribution of epidermis markers: K5 in the basal layer (white arrow); K14 in the basal layer (blue arrow); the marker of mature keratinocytes, K10 (red arrow); and Involucrin (pink arrow) in the upper layers.
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f3: Ultrastructural characteristics of hAFS cells and differentiation in 3D culture(a) Transmission electron microscope images of hAFS cells, hAFS-K and mature keratinocytes. Insets show tonofibrils in hAFS-K and keratinocytes (blue arrows), which are missing in hAFS cells. (b) H&E images of normal human skin and the epidermis constructed by hAFS-K demonstrate a well-organized, multilayer epithelium that contains all morphological layers (blue lines) and a collagen matrix in which resident fibroblasts (red line) play the role of the dermis. An epidermis constructed by keratinocytes was used as a positive control. (c) Immunohistochemistry of an organotypic epidermis constructed by hAFS-K showing a normal distribution of epidermis markers: K5 in the basal layer (white arrow); K14 in the basal layer (blue arrow); the marker of mature keratinocytes, K10 (red arrow); and Involucrin (pink arrow) in the upper layers.

Mentions: To further examine the biological function of hAFS-K, we compared the cellular structure of hAFS cells, hAFS-K cells and mature keratinocytes. Although hAFS cells have an expanded endoplasmic reticulum, a clearly visible nucleolus, and a high ratio of nucleus to cytoplasm (consistent with a high proliferation rate), they lack tonofibrils (organelles typical of keratinocytes). In contrast, mature keratinocytes contain multi-filamentary structures and conspicuous tonofibrils and show a low ratio of nucleus to cytoplasm. hAFS-K cells display features of keratinocyte precursor cells, with tonofibrils distributed in the cytoplasm, but these cells also have a prominent nucleolus and a high nucleus-to-cytoplasm ratio (Fig. 3a).


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)

Ultrastructural characteristics of hAFS cells and differentiation in 3D culture(a) Transmission electron microscope images of hAFS cells, hAFS-K and mature keratinocytes. Insets show tonofibrils in hAFS-K and keratinocytes (blue arrows), which are missing in hAFS cells. (b) H&E images of normal human skin and the epidermis constructed by hAFS-K demonstrate a well-organized, multilayer epithelium that contains all morphological layers (blue lines) and a collagen matrix in which resident fibroblasts (red line) play the role of the dermis. An epidermis constructed by keratinocytes was used as a positive control. (c) Immunohistochemistry of an organotypic epidermis constructed by hAFS-K showing a normal distribution of epidermis markers: K5 in the basal layer (white arrow); K14 in the basal layer (blue arrow); the marker of mature keratinocytes, K10 (red arrow); and Involucrin (pink arrow) in the upper layers.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4477371&req=5

f3: Ultrastructural characteristics of hAFS cells and differentiation in 3D culture(a) Transmission electron microscope images of hAFS cells, hAFS-K and mature keratinocytes. Insets show tonofibrils in hAFS-K and keratinocytes (blue arrows), which are missing in hAFS cells. (b) H&E images of normal human skin and the epidermis constructed by hAFS-K demonstrate a well-organized, multilayer epithelium that contains all morphological layers (blue lines) and a collagen matrix in which resident fibroblasts (red line) play the role of the dermis. An epidermis constructed by keratinocytes was used as a positive control. (c) Immunohistochemistry of an organotypic epidermis constructed by hAFS-K showing a normal distribution of epidermis markers: K5 in the basal layer (white arrow); K14 in the basal layer (blue arrow); the marker of mature keratinocytes, K10 (red arrow); and Involucrin (pink arrow) in the upper layers.
Mentions: To further examine the biological function of hAFS-K, we compared the cellular structure of hAFS cells, hAFS-K cells and mature keratinocytes. Although hAFS cells have an expanded endoplasmic reticulum, a clearly visible nucleolus, and a high ratio of nucleus to cytoplasm (consistent with a high proliferation rate), they lack tonofibrils (organelles typical of keratinocytes). In contrast, mature keratinocytes contain multi-filamentary structures and conspicuous tonofibrils and show a low ratio of nucleus to cytoplasm. hAFS-K cells display features of keratinocyte precursor cells, with tonofibrils distributed in the cytoplasm, but these cells also have a prominent nucleolus and a high nucleus-to-cytoplasm ratio (Fig. 3a).

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