A novel method to generate and culture human mast cells: Peripheral CD34+stem cell-derived mast cells (PSCMCs)

作者：Schmetzer Oliver^*; Valentin Patricia; Smorodchenko Anna; Domenis Rossana; Gri Giorgia; Siebenhaar Frank; Metz Martin; Maurer Marcus

来源：Journal of Immunological Methods, 2014, 413: 62-68.

DOI：10.1016/j.jim.2014.07.003

• Mast cell
• IgE
• Immunoglobulin E
• Differentiation
• Culture
• In vitro

The identification and characterization of human mast cell (MC) functions are hindered by the shortage of MC populations suitable for investigation. Here, we present a novel technique for generating large numbers of well differentiated and functional human MCs from peripheral stem cells (= peripheral stem cell-derived MCs, PSCMCs). Innovative and key features of this technique include 1) the use of stem cell concentrates, which are routinely discarded by blood banks, as the source of CD34 + stem cells, 2) cell culture in serum-free medium and 3) the addition of LDL as well as selected cytokines. In contrast to established and published protocols that use CD34 + or CD133 + progenitor cells from full blood, we used a pre-enriched cell population obtained from stem cell concentrates, which yielded up to 10(8) differentiated human MCs per batch after only three weeks of culture starting with 10(6) total CD34 + cells. The total purity on MCs (CD117 +, Fc epsilon R1 +) generated by this method varied between 55 and 90%, of which 4-20% were mature MCs that contain tryptase and chymase and show expression of Fc epsilon RI and CD117 in immunohistochemistry. PSCMCs showed robust histamine release in response to stimulation with anti-Fc epsilon R1 or IgE/anti-IgE, and increased proliferation and differentiation in response to IL-1 beta or IFN-gamma. Taken together, this new protocol of the generation of large numbers of human MCs provides for an innovative and suitable option to investigate the biology of human MCs.

• 出版日期2014-11