Lymph Node Model Pack

Long term cell culture under flow outside the CO₂ incubator

Incubator independent setup

Culture cells up to 16 days outside the CO₂ incubator

Compatible with K-frame microscopes

The Stage top incubator was designed for live imaging

Low flow rates

Get as low as 2 µl/min

Lymph node model for lung metastasis

The prognosis of lung cancer patients with lymph node metastasis is deem. A study has found that the 5-year DFS (disease free survival) and OS (overall survival) of patients with detected lymph node metastasis of non-small cell lung cancer were lower (21,4%) than the negative group (61,8%) [Gwóźdź et al.]. Data like these call for better lymph node models to understand the reason behind the outcome and eventually prevent it.

Developing complex models like this always brings researchers to the tough crossroads of choosing between the overly-complex and ethically-demanding use of animal models and the overly-simplistic and less-physiologically relevant in vitro models.

Now there’s an in-between. Culturing cell underflow improves the physiological relevance of fluid dynamic organs such as the lymph nodes while also allowing researchers to better isolate variables. In addition, the lymph node model is automatized for longterm cell culture outside the CO2 incubator, increasing its flexibility and portability and decreasing the manual labor and human error.
lymph node model setup schematics

Setup

Cell culture pump

Flow sensor (Galileo, MIC)

Stage top incubator

Reservoirs

Tubings and fittings

Microfluidic chip

User guide

Software (Galileo user interface)

The setup includes the cell culture pump, which maintains the gas concentration fed into it to pressurize the liquid. For example, if a premixed bottle of 5% CO2 is plugged into the pump, mimicking a conventional CO2 incubator, it ensures that the media does not lose buffering CO2 to the atmosphere, so pH remains optimal for long periods outside the CO2 incubator. The flow sensor ensures that the flow is stable and reliable at a range of flow rates, starting at 2 µl/min.

 

The stage top incubator maintains the temperature at 37°C, but allows you to leave your cells on top of the microscope stage and perform live cell imaging with higher quality.

 

Reservoirs, tubing and connectors are all commercially available, can be reused or disposable, and can be bought independently and sterile. And the system was conceived to work with any type of chip, regardless if it’s commercial or home-made.

Lymph node model for lung metastasis in action

Our partners from the Tumor-LN-oC project cultured Lewis Lung Carcinoma (LLC) cells under flow in the project-specific chip by using the lymph node model pack.  Initially the culture was terminated after 3 days to test the viability of the cells. Following the successful results of the 3-day culture, the experiment was repeated. The culture was viable for up to 16 days, as illustrated below.
 
lymph node model results by collaborator
The experiment was terminated due to the consumption of the cell culture medium, a limitation that can be circumvented by adding a recirculation bridge.

If you want to know more about organ-on-a-chip technology, have a look at this review!
References
  1. Gwóźdź, M. Pasieka-Lis, K. Kołodziej, J. Pankowski, R. Banaś, M. Wiłkojć, et al. Prognosis of patients with stages I and II non-small cell lung cancer with nodal micrometastases. Ann. Thorac. Surg., 105 (5) (2018), pp. 1551-1557.
  2. Featured image description: This image of mouse lymph node was captured using multiphoton microscopy. B lymphocytes (red) accumulate inside B-cell follicle, while GFP-expressing myeloid cells (green) surround it. Collagen fibers (blue) run along lymph node conduits, capillary network (pink) delivers oxygen and nutrients to the follicle.

    Caption: B-cell follicle inside a mouse lymph node

    Wikimedia Commons, author Olena12

Compatibility and Applications

Some biological applications of our lymph node model pack include:

brain cancer stem cells cancer cell migration

Cancer Invasion and Metastasis

Follow long-term cancer cell migration in real time on top of the microscope

✓ Keep ideal temperature conditions

✓ Follow Cancer Cell Migration in Real Time

✓ Adapted to Cultures Under Flow

Cancer cell migration

Cell-to-cell interactions 

Metastasis on chip

And many more! 

And the lymph node pack could also be adapted to other types of organs: 

Gut-on-a-chip

Gut-on-a-chip pack

Intestinal cells coculture under flow, mimicking the gut physiology

✓ All microfluidic pieces included, quick and easy assembly

✓ Dynamic culture conditions

✓ Advanced in viro/ex vivo

Gut-on-chip

small intestine villi inflammatory bowel disease

Inflammatory bowel disease model

Automatically collect important markers of IBD in a relevant in vitro model

✓ Uncover cytokine profile changes in time

✓ Mimic pathological conditions of IBD

✓ Tailor sample volume to your analysis

Inflammatory bowel disease model

Blood-brain barrier on a chip mouse cappillaries

Blood-brain Barrier on Chip

Plug-and-play instrument pack for long term BBB on a chip study

✓ Relevant microenvironment

✓ Automatized organ-on-chip perfusion

✓ Plug-and-play microfluidic platform

Blood-brain Barrier on Chip

liver-on-chip model mouse tissue

Liver-on-a-chip model

Mimic the liver microenvironment in long term experiments

✓ Improve your reproducibility with physiological culturing conditions

✓ Automated and controlled supply of nutrients in a stable flow

✓ Test different conditions at the same time

Liver-on-chip

lung-cells

Lung-on-a-chip model pack

Perform lung research in a physiologically relevant microenvironment

✓ Culture your lung cells in a physiological air-liquid interface

✓ Continuous and controlled supply of nutrients in a stable flow

✓ Stop losing your cell experiment due to clogging

Lung-on-a-chip

And many more! 

Microfluidic perfusion for dynamic cell culture

Technical specifications for the lymph node model pack

The lymph node model pack is composed of two main pieces of equipment: the cell culture pump and the stage top incubator.

Cell culture pump technical specifications

The cell culture pump can come with 1 to 6 channels. Each channel connects to a flow sensor with a feedback-loop between pressure and flow rate. Each channel has the following specifications (tested with a set pressure value of 2 bar):

Characteristics Specifications
Accuracy +/- 2.5 mbar
Air consumption few mL/min
Response time 140 ms
Settling time 2750 ms
Overshoot 0.12 mbar

Stage top incubator technical specifications

The Stage Top Incubator was designed to allow cell culture on top of the microscope stage for live cell imaging.

CharacteristicsSpecifications
Dimensions (mm)30.5 x 130 x 168 (h x w x l)
Base K- Frame3.5 x 110 x 160 (h x w x l)
Dimensions of internal usable space25 x 89 x 130 (h x w x l)
Dimensions of the bottom glass (ITO glass)1) 72 x 110 with a thickness of 1.1 mm

2) 50 x 25 with a thickness of 0.6mm

3) 50 x 22 with a thickness of 0.12 mm

Temperature rangeRoom Temperature to 70 °C
Temperature accuracy± 0,5 °C
External MaterialAluminum

The stage top incubator is controlled by a converter connected to software (both included). Also, it is compatible with 1/16″ and 1/32″ ID tubing.

microfluidic-cell-culture-system-converter
Pressure splitter, to be kept outside the incubator (left); and cell culture platform (right), which can be entirely placed inside the incubator.

Frequently asked questions

Which gasses can the cell culture pump use for the lymph node model?

The cell culture pump can use any non-corrosive gas mixture.

To guarantee the sterility of the used gasses, we advise adding a small disposable filter at the gas inlet of the reservoir to keep the sterile of your lymph node model.

No, the stage top Incubator allows gas exchange with the atmosphere.

The system works well with the range of 0-5 ml/min.

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Tumor-LN-oC logo

Funding and Support

The Tumor-LN-oC project helped develop this instrument. This project is funded by European Union’s H2020-NMBP-TR-IND-2020, grant agreement-no. 953234 (Tumor-LN-oC).

Products & Associated Accessories

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