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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

 

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

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.

 

Characteristics

Specifications

Dimensions (mm)

30.5 x 130 x 168 (h x w x l)

Base K- Frame

3.5 x 110 x 160 (h x w x l)

Dimensions of internal usable space

25 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 range

Room Temperature to 70 oC

Temperature accuracy

± 0,5 oC

External Material

Aluminum 

 

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.

Compatibility and Applications

Lymph node model applications

Some biological applications of our lymph node model pack include:

 

 

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

Microfluidic perfusion for dynamic cell culture
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.

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).

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