Journal: International Journal of Molecular Sciences

Article Title: Characterization of 3D Organotypic Culture of Mouse Adipose-Derived Stem Cells

doi: 10.3390/ijms25073931

Figure Lengend Snippet: Assessment of 3D organotypic ADSC differentiation potential. ( A ) Representative images of 3D organotypic ADSCs during adipogenic differentiation on the 14th day. The decrease in organoid size relative to the nondifferentiation control group is apparent. Scale bar size is 200 μm. ( B ) Oil Red O staining performed on the 14th day of adipogenic differentiation. Positive staining for Oil Red O is observed only in peripheral cells of the matrix. Scale bar size is 400 μm. ( C ) Relative mRNA expression levels of adipocyte markers LPL and adipoQ in 3D ADSCs compared to 2D ADSCs as a positive control. The expression of LPL and adipoQ mRNA in 3D ADSCs was not detected, in contrast to their expression in 2D ADSCs. The values are the means ± standard deviations ( n = 4).

Article Snippet: Adipogenic induction media (STEMCELL Technologies, Vancouver, Canada) were then added and replaced every 3 days.

Techniques: Control, Staining, Expressing, Positive Control

Journal: Journal of Tissue Engineering

Article Title: Therapeutic potential of adipose-derived mesenchymal stem cell exosomes in tissue-engineered bladders

doi: 10.1177/20417314211001545

Figure Lengend Snippet: Identification of Ad-MSCs (a–c) and AMEs (d–f): (a) characterization of the fourth generation of Ad-MSCs was performed by flow cytometry, and the Ad-MSC histogram revealed that after purification, the cells mostly expressed mesenchymal cell markers. Differentiation assays showed that Ad-MSCs could differentiate into adipocytes (b) and osteocytes (c), which were stained with Oil Red O and Von Kossa, respectively. Scale bar = 100 μm. (d) TEM of exosomes isolated from MSCM-cultured Ad-MSCs for 3 days with 10% exosome-depleted FBS. Scale bars = 100 nm. (e) NTA found that most of these vesicles ranged in diameter from 30 to 150 nm. (f) Western blotting was performed with Ad-MSCs (MSCs) and AMEs (Exos). The expression of CD9, CD63, TSG101, and calnexin was detected. Ad-MSCs: adipose-derived mesenchymal stem cells; AMEs: adipose-derived mesenchymal stem cell exosomes; NTA: nanoparticle tracking analysis; TEM: transmission electron microscopy.

Article Snippet: The medium was changed to mesenchymal stem cell adipogenic differentiation medium (MADM, ScienCell) and mesenchymal stem cell osteogenic differentiation medium (MODM, ScienCell) when the cells reached 70–80% confluency.

Techniques: Flow Cytometry, Purification, Staining, Isolation, Cell Culture, Western Blot, Expressing, Derivative Assay, Transmission Assay, Electron Microscopy

Journal: Plastic and reconstructive surgery

Article Title: Adipose Tissue Engineering from Human Adult Stem Cells: Clinical Implications in Plastic and Reconstructive Surgery

doi: 10.1097/01.prs.0000244840.80661.e7

Figure Lengend Snippet: Human mesenchymal stem cells and their adipogenic differentiation in monolayer culture. (Above, left) Phase-contrast image of human mesenchymal stem cells undergoing proliferation. Note the fibroblast-like appearance of human mesenchymal stem cells (cf. Alhadlaq and Mao, 2004). (Below, left) Negative Oil-Red O staining of human mesenchymal stem cells without adipogenic induction, as in above, left. (Above, right) After treatment with adipogenic supplements of dexamethasone, insulin, and isobutyl-methylxanthine, the same population of human mesenchymal stem cells as in above, left treated with adipogenic stimulating medium for 1 week showed different cell morphology and the presence of rounded extracellular matrix vacuoles. (Below, right) Positive Oil-Red O staining of lipid vacuoles (arrow) of human mesenchymal stem cells treated with adipogenic induction indicates that human mesenchymal stem cells had differentiated into adipogenic cells.

Article Snippet: Inducing Human Mesenchymal Stem Cells to Differentiate into Adipocytes First-passage human mesenchymal stem cells were induced to differentiate into adipocyte-like cells by exposure to 10% human mesenchymal stem cell adipogenic differentiation medium (StemCell Technologies) with 1% antibiotic-antimycotic (Gibco) for 1 week (in vitro observations) and 4 weeks (for in vivo implantation).

Techniques: Staining

Journal: Plastic and reconstructive surgery

Article Title: Adipose Tissue Engineering from Human Adult Stem Cells: Clinical Implications in Plastic and Reconstructive Surgery

doi: 10.1097/01.prs.0000244840.80661.e7

Figure Lengend Snippet: Harvest of adipogenic poly(ethylene glycol)-based hydrogel grafts from human mesenchymal stem cells and control groups after in vivo implantation in athymic mice. (Above) Representative cell-free poly(ethylene glycol)-based hydrogel construct (between arrows) showing retention of the original size (9-mm diameter). (Center) Representative poly(ethylene glycol)-based hydrogel construct encapsulating human mesenchymal stem cells without adipogenic differentiation (between arrows) showing that poly(ethyleneglycol)-based hydrogel adhered to surrounding host tissue and retained the original size (9-mm diameter). (Below) Representative poly(ethylene glycol)-based hydrogel construct (between arrows) encapsulating adipogenic cells derived from human mesenchymal stem cells showing its adhesion to surrounding host tissue and retention of the original size (9-mm diameter; greater magnification).

Article Snippet: Inducing Human Mesenchymal Stem Cells to Differentiate into Adipocytes First-passage human mesenchymal stem cells were induced to differentiate into adipocyte-like cells by exposure to 10% human mesenchymal stem cell adipogenic differentiation medium (StemCell Technologies) with 1% antibiotic-antimycotic (Gibco) for 1 week (in vitro observations) and 4 weeks (for in vivo implantation).

Techniques: Control, In Vivo, Construct, Derivative Assay

Journal: Plastic and reconstructive surgery

Article Title: Adipose Tissue Engineering from Human Adult Stem Cells: Clinical Implications in Plastic and Reconstructive Surgery

doi: 10.1097/01.prs.0000244840.80661.e7

Figure Lengend Snippet: Shape, dimensions, and photo-opaqueness of in vivo harvested poly(ethylene glycol)-based hydrogel construct encapsulating engineered adipose tissue from human mesenchymal stem cells and control groups. (Above, left) Plastic cap of a 1.5-ml microcentrifuge tube (9-mm diameter) used as a generic mold of the shape and dimensions for engineered adipose tissue. (Above, right) Harvested cell-free poly(ethylene glycol)-based hydrogel is largely transparent. (Below, left) Poly(ethylene glycol)-based hydrogel encapsulating human mesenchymal stem cells (without adipogenic differentiation) showing some photo-opacity. (Below, right) Poly(ethylene glycol)-based hydrogel encapsulating adipogenic cells derived from human mesenchymal stem cells showing substantial photo-opacity. All poly(ethylene glycol) grafts maintained the original shape and dimensions (cf. above, left).

Article Snippet: Inducing Human Mesenchymal Stem Cells to Differentiate into Adipocytes First-passage human mesenchymal stem cells were induced to differentiate into adipocyte-like cells by exposure to 10% human mesenchymal stem cell adipogenic differentiation medium (StemCell Technologies) with 1% antibiotic-antimycotic (Gibco) for 1 week (in vitro observations) and 4 weeks (for in vivo implantation).

Techniques: In Vivo, Construct, Control, Derivative Assay

Journal: Plastic and reconstructive surgery

Article Title: Adipose Tissue Engineering from Human Adult Stem Cells: Clinical Implications in Plastic and Reconstructive Surgery

doi: 10.1097/01.prs.0000244840.80661.e7

Figure Lengend Snippet: Representative hematoxylin and eosin and Oil-Red O staining of tissue-engineered poly(ethylene glycol)-based hydrogel grafts retrieved after 4-week in vivo implantation in the dorsum of athymic mice (hMSCs, human mesenchymal stem cells). (Above) Representative hematoxylin and eosin– and Oil-Red O–stained micrographs of cell-free control poly(ethylene glycol)-based hydrogels construct showing neither resident cells nor lipid vacuoles. (Center, left) Representative hematoxylin and eosin–stained micrograph of poly(ethylene glycol)-based hydrogel construct encapsulating human mesenchymal stem cells demonstrates abundant resident cells. (Center, right) Representative Oil-Red O–stained micrograph of poly(ethylene glycol)-based hydrogel construct encapsulating human mesenchymal stem cells demonstrates a lack of lipid vacuoles. (Below, left) Representative hematoxylin and eosin–stained micrograph of poly(ethylene glycol)-based hydrogel construct encapsulating human mesenchymal stem cell– derived adipogenic cells demonstrates abundant resident cells among irregular islands of space. (Below, right) Representative Oil-Red O–stained micrograph of poly-(ethylene glycol)-based hydrogel construct encapsulating human mesenchymal stem cell– derived adipogenic cells demonstrates abundant lipid vacuoles among flattened cells that resemble adipocytes. It is probable that lipid vacuoles occupied the irregular islands of space seen in below, left (original magnification, ×10).

Article Snippet: Inducing Human Mesenchymal Stem Cells to Differentiate into Adipocytes First-passage human mesenchymal stem cells were induced to differentiate into adipocyte-like cells by exposure to 10% human mesenchymal stem cell adipogenic differentiation medium (StemCell Technologies) with 1% antibiotic-antimycotic (Gibco) for 1 week (in vitro observations) and 4 weeks (for in vivo implantation).

Techniques: Staining, In Vivo, Control, Construct, Derivative Assay

Journal: NPJ Regenerative Medicine

Article Title: Personalized medicine for reconstruction of critical-size bone defects – a translational approach with customizable vascularized bone tissue

doi: 10.1038/s41536-021-00158-8

Figure Lengend Snippet: a EPC after uptake of acetylated LDL-DiI (red) and lectin staining (green). Counterstaining with DAPI (blue). Scale bar 50 µm. b CD34 immunofluorescence staining (green) of EPC. Counterstaining with DAPI (blue). Scale bar 50 µm. c Sprouting of EPC spheroids after 48 h incubation in a fibrin gel. Scale bar 50 µm. d mRNA expression of EPC compared to MSC (set to 1). e Oil Red O staining of MSC after adipogenic differentiation. Scale bar 100 µm. f Alcian Blue staining of MSC after chondrogenic differentiation. Scale bar 200 µm. g Alizarin Red staining of MSC after osteogenic differentiation. Scale bar 200 µm.

Article Snippet: For the adipogenic differentiation, MSC were cultivated in a differentiation medium consisting of MSC adipogenic differentiation basal medium, supplemented with FCS, glutamine, insulin, 3-isobutyl-1-methyl-xanthine (IBMX), dexamethasone, rosiglitazone, and penicillin-streptomycin; for chondrogenic differentiation MSC were cultivated in MSC chondrogenic differentiation basal medium, supplemented with FCS, dexamethasone, ascorbate, sodium selenite (ITS), proline, TGF-β3, and penicillin-streptomycin; for osteogenic differentiation MSC were cultivated in MSC osteogenic differentiation basal medium, supplemented with FCS, glutamine, ascorbate, β-glycerophosphate, dexamethasone, and penicillin-streptomycin according to the manufacturer’s recommendation (all from Pelobiotech GmbH, Planegg, Germany).

Techniques: Staining, Immunofluorescence, Incubation, Expressing

Journal: BMC Cancer

Article Title: ADSCs and adipocytes are the main producers in the autotaxin–lysophosphatidic acid axis of breast cancer and healthy mammary tissue in vitro

doi: 10.1186/s12885-018-5166-z

Figure Lengend Snippet: Different mammary cells respond to stimulation with LPA 18:1 but express a different LPAR profile: ( a / b ) Mean relative LPAR mRNA expression. Values are calculated using the 2 -ΔCq method, the reference is HPRT ; ( a ) LPAR mRNA profile of ADSCs, MES and HMEC/BCC from tumors and healthy tissue; ( b ) The effect of adipogenic differentiation (Diff) on the LPAR mRNA profile of ADSCs compared to the control (CTL). ( c ) The effect of LPA on cytosolic free calcium levels; x-axis shows the molarity of LPA 18:1 in a common logarithmic scale, y-axis shows the maximal ratio of the emission of Fura-2 (340 nm/380 nm excitation). The mean measuring points with the SD are plotted and connected with a nonlinear fit (variable slope) using GraphPad Prism 7.00, epithelial cells have a significantly higher calcium release than ADSCs and mesenchymal cells at concentrations of 1 μM LPA and above (not plotted for clarity, p < 0.0167, Kruskal–Wallis H test, Mann–Whitney U test with Bonferroni correction); abbreviations: healthy h, tumor-distant td, tumor-adjacent ta, tumor t, EpCAM-positive breast cancer cells BCC; n = 4

Article Snippet: Adipogenic differentiation of the ADSCs was achieved using a mesenchymal stem cell adipogenic differentiation medium (PELOBIOTECH GmbH, Planegg/Martinsried, Germany) according to the manufacturer’s protocol.

Techniques: Expressing, Control, MANN-WHITNEY