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Immortalized human kerationocyte cell line

A technology of keratinocytes and cell lines, applied in the field of infinitely proliferating human keratinocytes, can solve the problem of poor understanding of tumor molecular activities

Inactive Publication Date: 2001-12-12
WISCONSIN ALUMNI RES FOUND
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although tumor recurrence near the primary tumor site is a major cause of treatment failure and death in these patients, little is known about the molecular events that lead to tumor regrowth after treatment

Method used

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  • Immortalized human kerationocyte cell line
  • Immortalized human kerationocyte cell line
  • Immortalized human kerationocyte cell line

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0234] Treatment Adhesive Suspended Example 1 Parent (*4th generation) 1.51% 43.3%

Embodiment 2

[0235] BC-1-Ep / SL (31st generation) 1.32% 69.8% Example 2 Parental generation (6th generation) 1.79% 32.8%

[0236]BC-1-Ep / SL (33rd generation) 1.05% 73.7%

[0237] Table 4

[0238] Effect of DNA concentration on transient transfection efficiency in BC-1-Ep / SL cells

[0239] DNA concentration

8 / 26 / 97

%GFP + cells

12 / 16 / 97

%GFP + cells

12 / 22 / 97

%GFP + cells

15 micrograms pGL + 5 micrograms pcDNA3

3.79％

13.64％

9.88％

20 micrograms pGL + 6.7 micrograms pcDNA3

3.2％

5.25％

8.85％

25 micrograms pGL + 8.3 micrograms pcDNA3

1.98％

12.49％

3.33％

Cell confluence

70％

35-40％

35.40％

[0240] table 5

[0241] The ratio of DNA to GeneFECTOR affects transient transfection efficiency

[0242] The ratio of DNA to Genefector

1 / 30 / 98

%GFP + cells

2 / 20 / 98

%GFP + cells

2 / 20 / 98

%GFP + cells

1∶2

...

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PUM

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Abstract

A spontaneously immortalized human keratinocyte cell line is disclosed. In a preferred embodiment, this cell line is ATCC 12191. In another embodiment of the invention, a method of assaying the effect of a test tumor cell modulation agent is disclosed. The method comprises the steps of obtaining a human stratified squamous epithelial cell culture, wherein the culture comprises human malignant squamous epithelial cells and spontaneously immortalized human keratinocytes, wherein the culture forms a reconstituted epidermis. One then treats the epidermis with a test tumor cell modulation agent and evaluates the growth of the malignant cells within the epidermis.

Description

[0001] Cross-documentation of related applications [0002] Not applicable. [0003] Statement regarding research or development sponsored by the federal government [0004] The present invention was completed with NIH Grant No. AR40284 granted by the US government. The US government has certain rights in this invention. Background of the invention [0005] Human keratinocytes [0006] Human keratinocytes isolated from stratified squamous epithelium are easy to grow in vitro (for a review see Leigh, et al., 1994). Cultivated keratinocytes are easy to replicate during early passages and can produce a large number of cells, which show certain characteristics of differentiation in squamous cells. When cultured normal human keratinocytes are transplanted into mice, the epithelial tissue structure will regenerate in an orderly manner over time (Breitkreutz, et al., 1997). The cultivation and transplantation of human keratinocytes is easy, and...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61P35/00A61P43/00C12N1/00C12N5/071C12N5/10C12N15/09C12Q1/02G01N33/15G01N33/50
CPCC12N2503/02C12N2502/094C12N2503/06C12N2502/1323C12N2510/00C12N5/0629C12N2502/30C12N2510/04G01N33/5014G01N2500/10C12N5/0698A61P35/00A61P43/00C12N5/0602C12N5/0693
Inventor L·艾伦-霍夫曼S·J·施洛斯尔M·A·皮卡特
Owner WISCONSIN ALUMNI RES FOUND
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