Kemshead J T, Askonas B A
Immunology. 1978 Jun;34(6):1071-6.
Culture dishes precoated with thin layers of acid soluble rat tail collagen simplify conditions necessary to obtain in vitro high IgG anti-DNP responses from primed and boosted mice. In contrast to Mishell-Dutton type cultures the use of collagen precludes the need to continually rock cultures and to add nutrients. Reduction of the cell density in collagen dishes to 10(6) cells/ml is without detriment to the antibody response. Cells adhering to collagen coated dishes after 24 h incubation provide an enriched population of IgG anti-DNP precursor cells; which will mature to antibody secretion at very low cell density (10(4) -10(5) cells/ml). However, as changes in the adhesive properties of cells to collagen occur during cell maturation, attempts to adapt the technique to obtain clones of antibody secreting cells were not wholly successful.
预先用薄层酸溶性大鼠尾胶原包被的培养皿简化了从致敏和加强免疫的小鼠中获得体外高IgG抗DNP反应所需的条件。与米舍尔-达顿(Mishell-Dutton)型培养不同,使用胶原可避免持续摇动培养物和添加营养物质的需要。将胶原培养皿中的细胞密度降低到10⁶个细胞/毫升对抗体反应没有损害。孵育24小时后附着在胶原包被培养皿上的细胞提供了丰富的IgG抗DNP前体细胞群体;这些细胞将在非常低的细胞密度(10⁴ - 10⁵个细胞/毫升)下成熟并分泌抗体。然而,由于细胞成熟过程中细胞与胶原的粘附特性会发生变化,试图采用该技术获得抗体分泌细胞克隆并未完全成功。