Journal: Cellular and Molecular Life Sciences

Article Title: Bioengineered MSC Cxcr2 transdifferentiated keratinocyte-like cell-derived organoid potentiates skin regeneration through ERK1/2 and STAT3 signaling in diabetic wound

doi: 10.1007/s00018-023-05057-3

Figure Lengend Snippet: Cxcr2-mediated increased transdifferentiation potential of MSCs toward keratinocyte-like cells a Differential expression analysis of keratinocyte and epithelial markers in MSC GFP , MSC Cxcr2 , MSC Scr , and MSC Cxcr2 KD -derived KLCs revealed a significant increase in the expression of Basonuclin ( Bnc1 ), Cytokeratin 5, 14, 1, Involucrin ( Ivl ), Stratifin ( Sfn ), E-Cadherin ( Cdh1 ), and Mucin-1 ( Muc1 ) in MSC Cxcr2 -KLCs. b Confocal microscopy images of MSC GFP , MSC Cxcr2 , MSC Scr , and MSC Cxcr2 KD -derived KLCs immuno-stained with Involucrin and CK5 and c Involucrin and CK14 showing a marked increase in the expression of Involucrin, CK5, and CK14 in MSC Cxcr2 -derived KLCs. ( n = 3, * p < 0.05 as compared with MSC and MSC GFP -KLC)

Article Snippet: Transdifferentiation of Cxcr2-bioengineered MSCs into keratinocyte-like cells (KLCs): Cxcr2-bioengineered MSC trans-differentiation in 2D culture: Cxcr2 -bioengineered MSCs with stable overexpression and/or silencing were cultured for 14 days in a keratinocyte expansion medium containing keratinocyte growth supplement (HiMedia, Mumbai, India) for trans-differentiation into keratinocyte-like cells (KLCs).

Techniques: Quantitative Proteomics, Derivative Assay, Expressing, Confocal Microscopy, Staining

Journal: Cellular and Molecular Life Sciences

Article Title: Bioengineered MSC Cxcr2 transdifferentiated keratinocyte-like cell-derived organoid potentiates skin regeneration through ERK1/2 and STAT3 signaling in diabetic wound

doi: 10.1007/s00018-023-05057-3

Figure Lengend Snippet: Enhanced spheroid forming efficiency of MSC Cxcr2 -KLCs. a Representative bright-field microscopy images of Cxcr2 -modulated MSCs in spheroid culture assay at days 3, 7, and 14. b Graph depicting a significant increase in MSC Cxcr2 -KLC spheroid area (lower panel depicting the graph of linear regression analysis). c Differential expression analysis of keratinocyte markers in spheroids depicting a significant increase in the expression of Ck5 , Ck14 , Ck13 , Sfn , and Ivl . Confocal microscopy images of spheroids immuno-stained with d CK14, and Myc, e Involucrin, and CK5, depicting increased expression in MSC Cxcr2 -KLC spheroids. ( n = 3, * p < 0.05 as compared with MSC-KLC/MSC GFP -KLC/MSC Scr -KLC/MSC Cxcr2 KD -KLC; day 4/8 in spheroid area; and MSC in gene expression analysis)

Article Snippet: Transdifferentiation of Cxcr2-bioengineered MSCs into keratinocyte-like cells (KLCs): Cxcr2-bioengineered MSC trans-differentiation in 2D culture: Cxcr2 -bioengineered MSCs with stable overexpression and/or silencing were cultured for 14 days in a keratinocyte expansion medium containing keratinocyte growth supplement (HiMedia, Mumbai, India) for trans-differentiation into keratinocyte-like cells (KLCs).

Techniques: Microscopy, Quantitative Proteomics, Expressing, Confocal Microscopy, Staining, Gene Expression

Journal: Cellular and Molecular Life Sciences

Article Title: Bioengineered MSC Cxcr2 transdifferentiated keratinocyte-like cell-derived organoid potentiates skin regeneration through ERK1/2 and STAT3 signaling in diabetic wound

doi: 10.1007/s00018-023-05057-3

Figure Lengend Snippet: Enhanced re-epithelialization with MSC Cxcr2 transplantation at type 1 diabetic wound bed a Representative confocal microscopy images of regenerated type 1 diabetic mice wound tissue co-immunostained with CXCR2 and Myc tag, b showing significantly increased Pearson’s correlation coefficient in MSC Cxcr2 Tx group. c Representative confocal microscopy images of type 1 diabetic mice wound tissue co-immunostained with the keratinocyte markers Involucrin and CK14, d showing significantly increased Pearson’s correlation coefficient in MSC Cxcr2 Tx group ( n = 3, * p < 0.05 as compared with Un- Tx /MSC GFP Tx /MSC Scr Tx /MSC Cxcr2 KD Tx ) (Scale bar: 10×–100 µm, 60×–20 µm)

Article Snippet: Transdifferentiation of Cxcr2-bioengineered MSCs into keratinocyte-like cells (KLCs): Cxcr2-bioengineered MSC trans-differentiation in 2D culture: Cxcr2 -bioengineered MSCs with stable overexpression and/or silencing were cultured for 14 days in a keratinocyte expansion medium containing keratinocyte growth supplement (HiMedia, Mumbai, India) for trans-differentiation into keratinocyte-like cells (KLCs).

Techniques: Transplantation Assay, Confocal Microscopy

Journal: Cellular and Molecular Life Sciences

Article Title: Bioengineered MSC Cxcr2 transdifferentiated keratinocyte-like cell-derived organoid potentiates skin regeneration through ERK1/2 and STAT3 signaling in diabetic wound

doi: 10.1007/s00018-023-05057-3

Figure Lengend Snippet: Generation of bi-layered 3D skin organoids with fibroblasts and bioengineered MSC-KLCs a Schematic representation of the organoid culture system. b Differential expression analysis of keratinocyte and epithelial markers in MSC GFP -KLC (control) organoids and MSC Cxcr2 -KLC organoids depicting a significantly high expression of the keratinocyte markers Ck1 and Flg , and epithelial marker Cldn1 in MSC Cxcr2 -KLC organoids. c Representative H&E-stained images of the skin organoids. d Representative confocal microscopy images of whole mount MSC GFP -KLC and/or MSC Cxcr2 -KLC organoids, co-immunostained with Myc and CK5 (upper panel), and Involucrin and CK14 (lower panel). e Quantification of Pearson’s correlation coefficient depicting significant co-localization of Myc and CK5 (left), and Involucrin and CK14 (right) in MSC Cxcr2 -KLC organoids ( n = 3, * p < 0.05 as compared with MSC GFP -KLC organoids) (Scale bar: 60×–20 µm)

Article Snippet: Transdifferentiation of Cxcr2-bioengineered MSCs into keratinocyte-like cells (KLCs): Cxcr2-bioengineered MSC trans-differentiation in 2D culture: Cxcr2 -bioengineered MSCs with stable overexpression and/or silencing were cultured for 14 days in a keratinocyte expansion medium containing keratinocyte growth supplement (HiMedia, Mumbai, India) for trans-differentiation into keratinocyte-like cells (KLCs).

Techniques: Quantitative Proteomics, Control, Expressing, Marker, Staining, Confocal Microscopy