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Getting started with Cellix's Kima Pump


Introduction

Kima pump is a microfluidic pump designed to aid cell culture (e.g. endothelial cells) under flow in various biochips and flow chambers, including Cellix’s Vena8 Endothelial+ biochips where it is possible to culture 8 cell monolayers simultaneously over 24–48 hours. Cell culture can be continued for up to several weeks if required. The Kima pump is controlled by the iKima Application on the iPod touch. The Kima pump is placed inside a standard incubator while the iPod Touch, as it’s controller, sits outside the incubator. The Kima pump connects easily to Cellix’s Vena8 Endothelial+ biochip and other manufacturer’s flow chambers via fully autoclavable tubing and delivers pulses of fresh media to cells seeded inside the microchannels of the biochip. The iKima App is available for download from Apple’s iTunes store.

Features and Benefits :

  • Cell Culture under flow: simple culture of endothelial cell monolayers under flow in flow chambers and 8 cell monolayers in Cellix's Vena8 Endothelial+ biochips within 24-48 hours.

  • Flow Rates: ~250 - 600µL/min. If using Kima pump with Vena8 biochip range, the flow splitter will result in flow rates of ~30 - 75µL/min in each of the 8 channels of the biochip.

  • Recirculating Culture Media: saves reagents and no additional fluidic valves required.

  • Incubator Safe: No additional temperature or humidity control required.

  • Pulsatile: apply continuous or periodic pulses of fresh culture media to cell monolayers in culture.

  • Programmable: with iPod Touch.

  • Easy to Use: control up to 4 pumps with the same controller and iPod Touch.

Applications :

  • Cell culture of endothelial cell monolayers for shear flow experiments.​

  • Cell-cell adhesion shear flow assays on endothelial cell monolayers with Cellix's Mirus Evo or ExiGo pump for studies such as:

  1. Cardiovascular / Atherosclerosis​: primary monocyte adhesion to coronary artery endothelial cells.

  2. Inflammation​: THP-1 monocyte adhesion to HUVECs under flow.

  3. Asthma / COPD: eosinophil adhesion studies to human lung microvascular endothelial cells (HMVEC-L).

  4. Oncology - Breast Cancer: MDA cells adhering to TNF-alpha stimulated HUVECs under shear flow.

Kima Pump Perfusion Protocol: coating and cell seeding in Vena8 Endothelial+ biochips and perfusion using Kima pump

NOTE: Kima pump is only for exchange of culture media to feed the cells (i.e. pulses of fluid / pulsatile flow) rather than a continuous flow.

Step 1:

Cellix’s Vena8 Endothelial+ biochips are coated using a standard pipette tip. Dispense ~12 μL of protein (e.g. fibronectin) into each microchannel. Note the excess of liquid on the entrance and exit ports.


Step 2:

The Vena8 Endothelial+ biochip is then placed in a humidified sterile petri dish, which should be placed at 4°C for overnight coating.


Step 3:

(a) After the incubation period, add 5 μL of 1.5 x 106 /100 μL (≅ 15 x 106/mL)

of endothelial cells gently into each channel. *Note: concentration specified is for primary HUVEC.

(b) The biochip is kept in a sterile petri dish and in the CO2 incubator for 15–20 minutes. Observe the biochip under microscope and top up all the reservoirs with 50 μL of media. Keep the biochip for 1.5–2 hours in the CO2 incubator.

Step 4:

Wash Kima pump with 70% ethanol and then with sterile media using a 5 mL sterile syringe in the biosafety hood.


Step 5:

Connect tubing from media bottle to the inlet port of the pump. Connect tubing from outlet port of the pump (8-way cable with pins) to a sterile petri dish. Wash the pump using iKima app with media for 3 minutes.


Step 6:

(a) Take the biochip from the incubator and place in the biosafety hood. Before connecting to the biochip, start perfusion using iKima app — typically 2 minutes perfusion, followed by 15–20 minutes pause. When media droplets form at the pins, gently connect the 8-way cable to the biochip to avoid air bubbles.

(b) Connect the outlet pins to the biochip which is connected to a discard bottle or to the same media bottle for recirculation of the same media. This is done in the biosafety hood. Once connected, transfer Kima platform to a CO2 incubator.

Note: all tubing and bottles must be autoclaved prior to the experiment.


So now you know the features and benefits of the Kima Pump and how it works.

But, if you still have questions or want to learn more, check out our website, at wearecellix.com, or contact us at info@wearecellix.com.

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