primary human dermal lymphatic endothelial cells lecs  (PromoCell)

Journal: Materials Today Bio

Article Title: Three-dimensional modelling of lymphangiogenesis in-vitro using bioorthogonal click-crosslinked gelatin hydrogels

doi: 10.1016/j.mtbio.2025.102367

Figure Lengend Snippet: Three-dimensional (3D) distribution of HDLEC within GelTN hydrogels. (A) Orthogonal section view of Z-stack confocal images (700 μm) of HDLEC encapsulated in GelTN (6–12 % w/v), stained with calcein-AM, presented with depth coding to indicate the z-position (depth) of HDLEC within GelTN network. The scale bar indicates the depth range of 0–700 μm, as a reference for the z-axis positioning of HDLEC within GelTN hydrogels. (B) Schematic illustration of HDLEC encapsulation in GelTN hydrogel, followed by compatibility assessments. (C) Top view (X – Y plane) of z-stack confocal images of HDLEC embedded in GelTN (6–12 %) hydrogels for up to 14 days, stained with calcein AM (green) and propidium iodide (red). Scale bar = 200 μm.

Article Snippet: Primary human dermal lymphatic endothelial cells (HDLEC) isolated from single donor juvenile foreskin, were purchased from PromoCell® (Heidelberg, Germany).

Techniques: Staining, Encapsulation

Journal: Materials Today Bio

Article Title: Three-dimensional modelling of lymphangiogenesis in-vitro using bioorthogonal click-crosslinked gelatin hydrogels

doi: 10.1016/j.mtbio.2025.102367

Figure Lengend Snippet: Quantification of HDLEC network formation, viability and metabolic activity in response to 3D encapsulation in GelTN. (A) Percentage of viable cells after 24 h of encapsulation. (B) Quantification of (i) branches, (ii) isolated elements and (C) total network formation of HDLEC-laden in GelTN hydrogels (6–12 %) after 14 days. (D) Metabolic activity of HDLEC encapsulated in GelTN (6–12 %) at day 1, 5, 10 and 14, presented as relative fluorescence unit (RFU) of AlamarBlue. Data are presented as mean ± SD (n = 3).

Article Snippet: Primary human dermal lymphatic endothelial cells (HDLEC) isolated from single donor juvenile foreskin, were purchased from PromoCell® (Heidelberg, Germany).

Techniques: Activity Assay, Encapsulation, Isolation, Fluorescence

Journal: Materials Today Bio

Article Title: Three-dimensional modelling of lymphangiogenesis in-vitro using bioorthogonal click-crosslinked gelatin hydrogels

doi: 10.1016/j.mtbio.2025.102367

Figure Lengend Snippet: Sprouting of HDLEC spheroids in GelTN at different concentrations after 24 h of encapsulation . (A) Schematic illustration of HDLEC spheroid generation and encapsulation using hanging drop method, followed by spheroid encapsulation in GelTN. (B) Representative brightfield (BF) images of HDLEC spheroids (scale bar = 250 μm) and z-stack confocal images (projection of x – y planes) of HDLEC spheroids stained with Hoechst (Blue) and Phalloidin (actin filaments, red). Scale bar = 200 μm. (C) Schematic illustration of the quantification method of HDLEC spheroid sprouts. (D) The quantification of HDLEC spheroids (i) sprout length and (ii) number. ( E) Quantification of (i) actin intensity and (ii) nuclei count per sprout. The quantification was carried out on 6 random spheroids for each experimental group. Data are presented as mean ± SD (n = 3).

Article Snippet: Primary human dermal lymphatic endothelial cells (HDLEC) isolated from single donor juvenile foreskin, were purchased from PromoCell® (Heidelberg, Germany).

Techniques: Encapsulation, Staining

Journal: Materials Today Bio

Article Title: Three-dimensional modelling of lymphangiogenesis in-vitro using bioorthogonal click-crosslinked gelatin hydrogels

doi: 10.1016/j.mtbio.2025.102367

Figure Lengend Snippet: Sprouting of HDLEC spheroids encapsulated in GelTN_Lo (6 % w/v) and GelTN_Hi (12 % w/v) hydrogels in response to vascular endothelial growth factor C (VEGF-C). (A) BF images of HDLEC spheroids embedded in GelTN_Lo and GelTN_Hi, supplemented with complete media (CTRL), or serum starved media (0.1 % FCS) with different concentrations of VEGF-C (0, 50, 100, and 200 ng/mL). (B) Quantification of HDLEC spheroids sprout length (left) and number (right) was calculated for 6 randomly selected spheroids per experimental group. (C) Representative confocal images of HDLEC after 72 h of hydrogel encapsulation (nuclei = blue, actin = red; scale bar = 200 μm).

Article Snippet: Primary human dermal lymphatic endothelial cells (HDLEC) isolated from single donor juvenile foreskin, were purchased from PromoCell® (Heidelberg, Germany).

Techniques: Encapsulation

Journal: Materials Today Bio

Article Title: Three-dimensional modelling of lymphangiogenesis in-vitro using bioorthogonal click-crosslinked gelatin hydrogels

doi: 10.1016/j.mtbio.2025.102367

Figure Lengend Snippet: The effect of GelTN concentration on the hydrogel diffusion rate and mesh size and HDLEC on sprouting. GelTN_Lo and GelTN_Hi were prepared and incubated in 2 mg/mL FITC-Dextran (20 kDa) for up to 24 h with the (A) fluorescence measured post-digestion after 0, 2, 6 and 24 h, and (B) the percentage diffusion rate was calculated using the control hydrogel (containing 2 mg/mL FITC-Dextran). Statistical analysis represents the significant difference of diffusion rate between GelTN_Lo and GelTN_Hi. (C) Estimated mesh size of GelTN_Lo and GelTN_Hi (n = 3). (D) Bright field images of HDLEC spheroids encapsulated in: (Left) GelTN_Hi immersed in media containing 50 ng/mL VEGF-C, or (right) GelTN_Hi containing 50 ng/mL VEGF-C (GelTN_Hi_VC) for 24 h (scale bar = 100 μm). (E) The quantification of HDLEC sprout (i) number and (ii) length, embedded in GelTN_Hi and GelTN_Hi_VC compared to GelTN_Lo for 24 h.

Article Snippet: Primary human dermal lymphatic endothelial cells (HDLEC) isolated from single donor juvenile foreskin, were purchased from PromoCell® (Heidelberg, Germany).

Techniques: Concentration Assay, Diffusion-based Assay, Incubation, Fluorescence, Control, Hi-C

Journal: Materials Today Bio

Article Title: Three-dimensional modelling of lymphangiogenesis in-vitro using bioorthogonal click-crosslinked gelatin hydrogels

doi: 10.1016/j.mtbio.2025.102367

Figure Lengend Snippet: Mechanical regulation of HDLEC lymphangiogenic activity in GelTN_Lo and GelTN_Hi via matrix metalloproteases (MMPs) and integrins. (A) RT-qPCR of the relative expression of (i) MMP2 , MMP9 and MMP14 in HDLEC encapsulated in GelTN_Lo vs GelTN_Hi (n = 3; scale bar = 250 μm). (B) Brightfield images of HDLEC spheroids 24 h post embedding in GelTN_Lo supplemented with 50 ng/mL VEGF-C in the presence or absence of 50 μM Marimastat (MMPi). (C) Relative expression of α5 ( ITGα5 ), β1 ( ITGβ1 ), and β3 ( ITGβ3 ) integrins in HDLEC encapsulated in 1GelTN_Lo vs GelTN_Hi (n = 3). (D) Brightfield images of siCtrl and siβ3 transfected HDLEC spheroids encapsulated in GelTN_Lo, cultured in 50 ng/mL VEGF-C containing media (i and ii), or 50 ng/mL VEGF-C containing media with 20 μM K34C (iii and iv). Scale bar = 100–250 μm. (E) Quantification of HDLEC spheroid sprouts length (left) and number (right) was calculated for 9 randomly selected spheroids per experimental group. Data are presented as mean ± SD.

Article Snippet: Primary human dermal lymphatic endothelial cells (HDLEC) isolated from single donor juvenile foreskin, were purchased from PromoCell® (Heidelberg, Germany).

Techniques: Activity Assay, Quantitative RT-PCR, Expressing, Transfection, Cell Culture

Journal: Materials Today Bio

Article Title: Three-dimensional modelling of lymphangiogenesis in-vitro using bioorthogonal click-crosslinked gelatin hydrogels

doi: 10.1016/j.mtbio.2025.102367

Figure Lengend Snippet: Evaluation of GelTN hydrogel injectability and its potential as a growth factor delivery platform. (A) Schematic illustration of the mechanical test setup to measure the injection force of GelTN primed in 1 mL syringe and 26G needle. (B) Representative Force – Time plots of GelTN (6 % w/v), ejected at 0 then every 5 min at 20 min post-crosslinking up to 60 min (n = 5). Water was used as a control. (C) VEGF-C release from GelTN in comparison to VEGF-C in PBS (control; n = 3). (D) Percentage VEGF-C release determined by cumulative VEGF-C concentration divided by the initial total VEGF-C concentration, multiplied by 100 (n = 3). (E) Representative images of western blot analysis showing p-Akt (60 kDa) and p-ERK1/2 (44 and 42 kDA) levels in HDLEC treated with GelTN_VC, V_100, CM, SM and GF-free GelTN (NC), for 20- and 30-min. Total Akt and ERK1/2 levels measured in comparison to the phosphorylation levels. GAPDH (36 kDa) was used as a loading control (Left panel). Quantification of p-Akt/total-Akt ratio in HDLEC treated with supernatants from SM, GelTN, GelTN_VC, VC100 and CM, for 20 and 30 min (right panel, n = 4). (F) Metabolic activity of HDLEC after treatment with supernatants from GelTN_V hydrogels, GelTN hydrogels, and controls: CM, SM and 100 ng/mL VEGF-C (V_100) for 72 h. Percentage HDLEC metabolic activity relative to the SM control (n = 3). (G) Representative brightfield images of HDLEC spheroids encapsulated in GelTN and treated with SM, VC100 and GelTN_VC for 24 h (Scale bar = 200 μm). Data are presented as mean ± SD.

Article Snippet: Primary human dermal lymphatic endothelial cells (HDLEC) isolated from single donor juvenile foreskin, were purchased from PromoCell® (Heidelberg, Germany).

Techniques: Injection, Control, Comparison, Concentration Assay, Western Blot, Phospho-proteomics, Activity Assay