Patent ID: 11946071
Assignee: UNIVERSITÄT ZÜRICH
Field: Measurement (Instruments)
Classification: CPC C  G | IPC C  G

Claim 7:
8. A method for the generation of the engineered layered cellular aggregate of claim 1, comprising the steps of;
a) providing a population of myeloid cells, wherein said myeloid cells are provided
(i) ex-vivo from a patient by isolation from fresh blood using a double gradient centrifugation; or
(ii) ex vivo as cell culture or cell line characterized by expression of monocyte/macrophage cell markers;

b) in a differentiation-priming step, contacting said myeloid cells with a protein kinase C agonist, yielding primed myeloid cells;
c) in a culture step, incubating said primed myeloid cells in the presence of low-density lipoprotein (LDL) in a confined volume in a hanging drop culture; yielding a 3-dimensional culture, a sphere of myeloid cells; and
d) in a co-culture step, incubating said 3-dimensional culture of myeloid cells together with fibroblasts in a confined volume, in a hanging drop culture in the presence of LDL, yielding said layered cellular aggregate.