Different types of cell lines in 3D cell culture on LifeGel®

Human cell lines

PANC-1

pancreatic cancer

On the photos below there is results of Panc-1 triple staining designed for staining different cellular organels – DiO (green) for cell membranes, MitoLite (red) for mitochondria and Hoechst (blue) for nuclei.

 

MiaPaca-2

pancreatic carcinoma

Cells from photo on the left and in the middle have been cultured on LifeGel 48-well plates for 10-14 days, 2500 cells per well have been seeded. On the photo on the right, cells were cultured for 10 days on 96-well plate, they have been seeded 1000 cells per well.

CFPAC-1

pancreatic ductal adenocarcinoma

CFPAC-1 cells have been cultured on 96-well plate with LifeGel for 14 days.

LoVo

colorectal adenocarcinoma; derived from metastatic site

LS180

colorectal adenocarcinoma; derived from metastatic site

RKO

colon carcinoma

RKO cells have been cultured on 96-well LifeGel plates for 12 days, they have been seeded 1500 cells/well.

RCC

renal cell carcinoma

Cells have been fixed on LifeGel and fluorescently stained with antibodies for: F-actin (green colour), α-tubulin (red colour), nucleus (blue colour). Photos in the second line is with brightfield and darkfield.

Project have been discovered in cooperation with IBIDI. Photos made by Shokoufeh Teymouri, µ-Slide Angiogenesis, IBIDI GmbH.

MDA-MB-231

breast cancer

MDA-MB-231 cells have been cultured on LifeGel for 10 days.

PDX (Patient Derived Xenograft)

primary cells

3D ex vivo culture of patient-derived xenograft model of colorectal cancer. Tumor sample was dissociated and cultured on 96-well plate with LifeGel for 14 days. Microscope images were taken using ZEISS Telaval 31 Inverted Microscope at 10x objective.

U-2 OS

bone carcinoma

Photos below show U-2 OS cells with stable transfection of GFP growing on LifeGel. Cells have been cultured on 96‑well plate for 21 days, 500 cells per well was seeded (obj. 2,5x lens 1,6x).

 

HUVEC

Human umbilical vein endothelial cells for angiogenesis research

HEK 293

human embryonic kidney 293 cells

HEK 293 cells grown on LifeGel was triple stained with DiO for cell membranes (green colour), MitoLite for mitochondria (red colour) and Hoechst for nuclei (blue colour). Photos: obj. 10x lens 2,5x.

MSC

mesenchymal stem cells

Mesenchymal stem cells culture on LifeGel have been stained for actin (green), tubulin (red) and nuclei (blue).

Cells were fixed using 4% formaldehyde solution (incubation 10-30 minutes), then perforated with 0,1% Triton-X in 10% FBS (in PBS). Actin was stained using DY-490-Phalloidin from Dynomics (1:500 in PBS). Tubulin was stained using primary (Monoclonal anti-alpha-Tubulin antibody, mouse; 1:500) and secondary (Anti-mouse IgG-Atto594; 5ug/mL) antibodies. Nuclei were stained with Hoechst 33342 (10 ug/mL). Photos: obj. 10x lens 1x.

Murine cell lines

Pan_02

pancreatic ductal adenocarcinoma

.

Pan_02 cells 3D structures immunofluorescence staining of actin (green), tubulin (red) and nuclei (blue) growing on LifeGel.

Cells were fixed using 4% formaldehyde solution (incubation 10-30 minutes), then perforated with 0,1% Triton-X in 10% FBS (in PBS). Actin was stained using DY-490-Phalloidin from Dynomics (1:500 in PBS). Tubulin was stained using primary (Monoclonal anti-alpha-Tubulin antibody, mouse; 1:500) and secondary (Anti-mouse IgG-Atto594; 5ug/mL) antibodies. Nuclei were stained with Hoechst 33342 (10 ug/mL). Photos: obj. 10x lens 1x.

4T1

breast cancer

4T1 cells shown below have been cultured on 96-well LifeGel plate for 7 days. Cells were seeded 500 cells/well.

4T1 cells shown below have been cultured on µ-Slide angiogenesis plate with LifeGel for 7 days. Cells were seeded 500 cells/well.

On the photos below there is staining with cultured 4T1 cells on 96-well LifeGel plate. DiO was for cell membranes (green), MitoLite for mitochondria (red), Hoechst for nuclei (blue). Obj. 2x, lens 1x.

B16-F10

skin cancer, melanoma

Hamster cell lines

CHO

Chinese hamster ovary cells

Co-culture

Co-culture CHO cells with Jurkat T

CHO – Chinese hamster ovary cells
Jurkat T – human lymphocytes, leukemia