Saito Hirohisa, Kato Atsushi, Matsumoto Kenji, Okayama Yoshimichi
Department of Allergy and Immunology, National Research Institute for Child Health and Development, 2-10-1 Okura, Setagaya-ku 157-8535, Tokyo, Japan.
Nat Protoc. 2006;1(4):2178-83. doi: 10.1038/nprot.2006.344.
This protocol details a method for obtaining a considerable number of human mast cells from peripheral blood cells from normal donors without using stem cell mobilization treatment. By using the magnetic cell sorting system, 10(4)-10(5) cells are retrieved in the CD34+ fraction when 100 ml of blood is drawn from a healthy donor. When these cells are cultured using methylcellulose medium supplemented with stem cell factor and interleukin 6, 10(3)-10(4) mast cell colonies are formed in 6 weeks. The total mast cell number at 6 weeks of culture will be 10(6)-10(7).
本方案详细介绍了一种无需使用干细胞动员治疗即可从正常供体的外周血细胞中获取大量人肥大细胞的方法。通过使用磁性细胞分选系统,从健康供体抽取100毫升血液时,在CD34+组分中可获得10⁴-10⁵个细胞。当使用添加干细胞因子和白细胞介素6的甲基纤维素培养基培养这些细胞时,6周内可形成10³-10⁴个肥大细胞集落。培养6周时肥大细胞总数将达到10⁶-10⁷个。