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Co-cultures of multiple cell types mimic pulmonary cell communication in response to urban PM₁₀

¹ Laboratory of Pneumology, Lung Toxicology Unit, ³ Center for Molecular and Vascular Biology, Katholieke Universiteit Leuven, Leuven, Belgium, ² Subdirección de Investigación Básica, Instituto Nacional de Cancerología, Mexico City, Mexico.

CORRESPONDENCE: E. Alfaro-Moreno, Katholieke Universiteit Leuven, Laboratory of Pneumology, Lung Toxicology Unit, Herestraat 49, O&N1 bus-706, B-3000, Leuven, Belgium. Fax: 32 16347124. E-mail: ealfaro{at}salud.gob.mx

Keywords: Co-culture, cytokine profile, particles with a 50% cut-off aerodynamic diameter of 10µm, tricultures

Received: March 21, 2008
Accepted July 7, 2008

The current authors evaluated whether a system of co-cultures of relevant cells (pneumocytes (A549), macrophages (THP-1), mast cells (HMC-1) and endothelial cells (EAHY926)) would mimic the responses to particles with a 50% cut-off aerodynamic diameter of 10 µm (PM₁₀) previously reported in vivo. The role of mast cells was considered of special interest.

Single cultures, bicultures (A549 + HMC-1 in a 10:1 ratio; THP-1 + HMC-1 in a 2:1 ratio) and tricultures (A549 + THP-1 + HMC-1 in a 10:2:1 ratio) were exposed to urban PM₁₀ (24 h at 0, 10, 30 or 100 µg·cm^–2). Additionally, EAHY926 cells were introduced in inserts above the tricultures. The released cytokines were evaluated with a fluorescence-activated cell sorter array system.

THP-1 + HMC-1 bicultures and the tricultures released more granulocyte colony-stimulating factor (G-CSF), macrophage inflammatory protein (MIP)-1β, interleukin (IL)-1β, IL-8, IL-6, tumour necrosis factor- and MIP-1 in response to PM₁₀ than the sum of the single cultures. Tricultures with EAHY926 released more G-CSF, MIP-1, IL-8 and MIP-1β than the EAHY926 single culture.

The bicultures, tricultures and tricultures with EAHY926 provide results that are consistent with the local and systemic effects previously described for particulate matter effects, i.e. inflammation, endothelial dysfunction and bone marrow cell mobilisation. Mast cells seem to play a significant role in the co-culture responses.

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