mucin stain  (Vector Laboratories)

Journal: Scientific reports

Article Title: De novo reconstruction of a functional in vivo-like equine endometrium using collagen-based tissue engineering.

doi: 10.1038/s41598-024-59471-z

Figure Lengend Snippet: Figure 2. Characterization of eMSCs (a) and eECs (b). (A) The outgrowth of the mix population from the explanted tissue under MSC medium condition. (B) The eMSCs had a spindle-shaped morphology, and (D) positive Vimentin staining indicated the presence of this intermediate filament protein (red: nuclei in blue). (C) MSC-specific markers (CD29, 40, 90) were demonstrated by gene expression analysis. (E) As indicators of osteogenic potential, eMSCs exhibited positive ALP staining, and (F) positive alizarin red staining for extracellular calcium deposits under osteogenic culture conditions, confirming differentiation into osteogenic lineages. (G) Oil Red O staining emphasized the presence of intracellular lipid droplets, confirming adipogenic differentiation capability of the eMSCs. The characterization of eECs (b) revealed (H,K) The outgrowth of the epithelial cells from the explanted tissue under FBS-EC and KSR-EC medium condition respectively (I,N) a polygonal cell shape in both EC media tested (FBS-EC and KSR-EC, respectively). (J,O) Positive Pan Cytokeratin staining of the cytoskeleton was evident in eECs in both media. In FBS-EC media, eEC purity, determined by flow cytometry, was 94.78% (K), whereas in KSR media, purity reached 97.70% (P). (L,Q) PCR validated the gene expression for the eEC-specific marker Muc1 in eECs derived from both EC media. (Uncropped conventional PCR gels were shown in the Supplementary Uncropped Fig. 1A–F for eMSCs and Figure for eECs).

Article Snippet: Additionally, the secretory function of the endometrial glandlike structures was assessed by staining for MUC1 (sc-7313, Santa Cruz Biotechnology), with a protocol adapted from Piña et al.31.

Techniques: Staining, Gene Expression, Flow Cytometry, Marker, Derivative Assay

Journal: Scientific reports

Article Title: De novo reconstruction of a functional in vivo-like equine endometrium using collagen-based tissue engineering.

doi: 10.1038/s41598-024-59471-z

Figure Lengend Snippet: Figure 4. Morphology of eECs from passage 6 in; (A) KSR-EC medium, and (B) FBS-EC medium. (C) Muc1 gene expression was not detected in eECs cultured in KSR-EC medium. (D) In contrast, eECs cultured in FBS-EC medium did show Muc1 expression. Scale bar: 300 µm. (Uncropped conventional PCR gels were shown in the Supplementary Fig. 2).

Article Snippet: Additionally, the secretory function of the endometrial glandlike structures was assessed by staining for MUC1 (sc-7313, Santa Cruz Biotechnology), with a protocol adapted from Piña et al.31.

Techniques: Gene Expression, Cell Culture, Expressing

Journal: Scientific reports

Article Title: De novo reconstruction of a functional in vivo-like equine endometrium using collagen-based tissue engineering.

doi: 10.1038/s41598-024-59471-z

Figure Lengend Snippet: Figure 6. (A) Expression of the FOXA2 gene in in vivo endometrial tissue and in the in vitro 3D-ET. FOXA2 protein was localized to the nucleus of the in vitro gland-like structures. (B) Muc1 gene expression in in vitro 3D-ET and (C) in vivo tissue with MUC1 protein expression in the cytoplasm of the cells lining the gland. (Uncropped conventional PCR gels were shown in the Supplementary Fig. 5).

Article Snippet: Additionally, the secretory function of the endometrial glandlike structures was assessed by staining for MUC1 (sc-7313, Santa Cruz Biotechnology), with a protocol adapted from Piña et al.31.

Techniques: Expressing, In Vivo, In Vitro, Gene Expression