Organotypic intestinal culture and methods of use thereof
First Claim
1. An organotypic culture comprising an artificial stroma comprising a mixture of collagen and human fibroblasts isolated from a human colon or intestine, said stroma overlayed with epithelial cells isolated from fetal human colon, wherein said culture contains a growth factor that binds the insulin growth factor-1 (IGF-1) receptor selected from insulin and IGF-1, a growth factor that binds the epidermal growth factor (EGF) receptor selected from EGF and tumor growth factor-alpha (TGF-alpha), and the growth factor leukemia inhibitory factor (LIF), which binds the LIF receptor.
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Abstract
An organotypic culture comprises an artificial stroma overlayed with epithelial cells isolated from a human colon or intestine. The stroma comprises a mixture of collagen and human fibroblasts isolated from a human colon or intestine. The culture contains a factor that binds the IGF-1 receptor, a factor that binds the EGF receptor, and a factor that binds the LIF receptor. These factors may be added exogenously to the culture via medium or may be expressed by various recombinantly engineered cell types in the culture. The organotypic culture can result in growth that is in situ-like or emphasizes other physiological or morphological states, depending on the balance of factors in the growth media. The organotypic culture may be used in methods for screening of therapeutic, carcinogenic, or growth enhancement factors, or for treating intestinal injuries by applying to the site of an injury the intact culture or the components thereof.
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Citations
19 Claims
- 1. An organotypic culture comprising an artificial stroma comprising a mixture of collagen and human fibroblasts isolated from a human colon or intestine, said stroma overlayed with epithelial cells isolated from fetal human colon, wherein said culture contains a growth factor that binds the insulin growth factor-1 (IGF-1) receptor selected from insulin and IGF-1, a growth factor that binds the epidermal growth factor (EGF) receptor selected from EGF and tumor growth factor-alpha (TGF-alpha), and the growth factor leukemia inhibitory factor (LIF), which binds the LIF receptor.
- 8. An organotypic culture comprising an artificial stroma comprising a mixture of collagen and human fibroblasts isolated from a human colon or intestine, said stroma overlayed with epithelial cells isolated from fetal human colon, wherein said culture contains a growth factor that binds the IGF-1 receptor selected from insulin and IGF-1, a growth factor that binds the EGF receptor selected from EGF and tumor growth factor-alpha (TGF-alpha), the growth factor LIF, which binds the LIF receptor, the growth factor endothelin-3 (ET-3), which binds the ET-3 receptor, a growth factor that binds the hepatocyte growth factor (HGF) receptor selected from HGF and tumor growth factor-beta (TGF-beta), the growth factor stem cell factor (SCF), which binds the SCF receptor, and the growth factor autocrine motility factor (AMF), which binds the AMF receptor.
Specification