Comparison of short term in vitro cultured human mast cells from different progenitors - Peripheral blood-derived progenitors generate highly mature and functional mast cells.
During the last two decades different scientific groups have investigated the phenotype and function of in vitro generated human mast cells (MC). The cells have been shown to display variable surface markers and functional characteristics. The phenotypic differences may reflect different culture conditions, protocols or the use of different progenitors. To investigate the significance of different progenitors, we have compared MC generated from CD133(+) progenitor cells from cord blood (CBMC) or peripheral blood (PBMC). The progenitors were cultured for 7 weeks in the presence of IL-6 and SCF, with addition of IL-3 the first 3 weeks, and FCS during week 7. The phenotype of the established MC was characterized by surface marker expression levels, metachromasia, histamine and tryptase contents and their function was evaluated by receptor-mediated release of histamine and PGD(2). The generated metachromatic (<99%) MC were 75% tryptase(+), regardless of the source of progenitor cell. Expression of c-kit/CD117, CD203c, and FcepsilonRI was comparable. The density of c-kit/CD117 receptors on CBMC was higher that of PBMC (p<0.001). The density of CD203c and FcepsilonRI was higher on PBMC (p<0.001). PBMC contained more histamine (p<0.001), expressed more FcepsilonRI (p<0.001) and released more histamine (p<0.001) and PGD(2) (p<0.001) upon ligation of FcepsilonRI, than CBMC. Culture with IL-4 increased expression of tryptase, FcepsilonRI, CD117 and CD203c, secretion of histamine and PGD(2) of PBMC, and histamine secretion of CBMC. Cord and peripheral blood may give rise to different types of MC. The question addressed should determine the progenitor cell and protocol to be used.
Department of Paediatrics, Aarhus University Hospital, DK-8200 Aarhus N, Denmark.
Hanne Busk
HB
Andersen
Mette
M
Holm
Thea Eline
TE
Hetland
Christine
C
Dahl
Steffen
S
Junker
Peter Oluf
PO
Schiøtz
Hans Jürgen
HJ
Hoffmann
D018929:Cell Culture Techniques / Q000379:methods*; D002454:Cell Differentiation; D049109:Cell Proliferation; D002478:Cells, Cultured; D005312:Fetal Blood / Q000166:cytology; D006412:Hematopoietic Stem Cells / Q000166:cytology / Q000502:physiology*; D006636:Histamine Release; D006801:Humans; D015847:Interleukin-4 / Q000494:pharmacology; D008407:Mast Cells / Q000166:cytology / Q000502:physiology*; D010727:Phosphoric Diester Hydrolases / Q000378:metabolism; D015230:Prostaglandin D2 / Q000378:metabolism; D019009:Proto-Oncogene Proteins c-kit / Q000378:metabolism; D011755:Pyrophosphatases / Q000378:metabolism; D017455:Receptors, IgE / Q000378:metabolism; D053802:Tryptases / Q000378:metabolism
C427944:ENPP3 protein, human; C508598:IL4 protein, human; D017455:Receptors, IgE; D015847:Interleukin-4; D019009:Proto-Oncogene Proteins c-kit; D010727:Phosphoric Diester Hydrolases; D053802:Tryptases; D011755:Pyrophosphatases; D015230:Prostaglandin D2