Novel hepatocyte-like cells and hepatoblast-like cells derived from hbs cells

A cell, cell population technology, applied in animal cells, vertebrate cells, artificial cell constructs, etc., can solve problems such as uninstructed CYPs

Inactive Publication Date: 2009-07-29
CELLECTIS SA
View PDF7 Cites 4 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, this application does not teach anything about the activity of these most important CYPs

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • Novel hepatocyte-like cells and hepatoblast-like cells derived from hbs cells
  • Novel hepatocyte-like cells and hepatoblast-like cells derived from hbs cells
  • Novel hepatocyte-like cells and hepatoblast-like cells derived from hbs cells

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0329] raw material

[0330] The starting material for use in the present invention is suitably multipotent undifferentiated hBS cells, eg undifferentiated hBS cell lines. Such materials are available from Cellartis AB and are also available through the NIH Stem Cell Registry http: / / stemcells.nih.gov / research / registry / . Cellartis AB has 2 hBS cell lines (SA001 and SA002) and a subclone of SA002 (SA002.5) available through the NIH. In addition, 20 Cellartis cell lines are listed in the UK Stem Cell Bank. These hBS cell lines and additional SA167 and SA348 from Cellartis AB have been frequently used in the present invention. hBS cells recommended as starting material are characterized by the following: positive for alkaline phosphatase, SSEA-3, SSEA-4, TRA 1-60, TRA 1-81, Oct-4, SSEA-1, telomerase Activity and pluripotency in vitro and in vivo (the latter shown by telomere formation in immunodeficient mice) were negative (see figure 1 ) (method and protocol as previously sh...

Embodiment 2

[0335] Protocol for obtaining hepatocyte-like cells

[0336] Intrinsic factor protocol (induced by exposure to intrinsic factor secreted into the medium) Differentiation

[0337] a) hBS cells grown in mEF cell layer in IVF dishes (Falcon) were subjected to 37°C, 5% CO 2 and 95% humidity for up to 40 days to obtain hepatocyte-like cells. Media used (VitroHES with added 4 ng / ml human recombinant bFGF [Invitrogen] TM [Vitrolife AB]) is replaced every 7-21 days, usually every 14 days by discarding about 1-2 ml of old medium and adding 1-2 ml of fresh medium. After 18-30 days, use a sharp microcapillary or Stem Cell ToolTM (Vitrolife AB) as a cutting and transfer tool to isolate hepatocyte-like cells from culture, and subsequently pool the cells for long-term storage (freezing) or use immediately, or alternatively directly fix and stain in culture dishes or use as live cells. Cells are used, for example, in Cyp activity assays. mEFs appear to provide an important signal to supp...

Embodiment 3

[0351] Characterization with liver markers (see figure 2 , 3 and 4)

[0352] Hepatocyte-like cells exhibit the general morphology of hepatocytes, ie they have a polygonal cell shape, large cell diameter (approximately 25-50 [mu]M), are often binucleated and tend to accumulate lipid particles. Furthermore, they express several markers described for hepatocyte types, such as albumin, α1-antitrypsin, LFABP, CK18 and HNF3β. They no longer express the stem cell marker for undifferentiated cells, Oct-4. Certain presumably less mature hepatocyte-like cells still express the fetal liver marker AFP. These cells were preferentially found within colonies of differentiated hBS cells. DAPI (4',6'-diamidino-2-phenylindole hydrochloride, Sigma Aldrich) was used as a control to visualize nuclei. (About Matrigel TM CK18 expression in differentiated hepatocyte-like cells, see Figure 22 . )

[0353] For identification of proliferating hepatoblast-like progenitor cells, AFP, HNF4, CK19...

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to view more

PUM

PropertyMeasurementUnit
diameteraaaaaaaaaa
diameteraaaaaaaaaa
Login to view more

Abstract

The present invention relates to a novel hepatocyte-like cell population derived from hBS cells and to the potential use of such heopatocyte-like cells in e.g. medical treatment, drug screening and toxicity testing. Furthermore, the invention relates to hepatoblast-like cells that may have suitable characteristics so that they can be used for the same applications as the hepatocyte-like cells and that furthermore may be used in in vitro studies of hepatogenesis such as early hepatogenesis or hepato-regenerative disorders. Both the hepatocyte-like and the hepatoblast-like cells according to the invention express drug transporter and/or drug metabolising characteristics either at the gene or protein expression level.

Description

Background of the invention [0001] The present invention relates to novel populations of hepatocyte-like cells derived from hBS cells and the potential use of such hepatocyte-like cells in, for example, drug therapy, drug screening and toxicity testing. Furthermore, the present invention relates to hepatoblast-like cells that can have suitable characteristics so that they can be used for expansion and further differentiation into functional hepatocyte-like cells if desired, and further can be used for in vitro and in vivo hepatogenic studies, such as early hepatogenesis or disorders of liver regeneration. The hepatocyte-like cells according to the present invention express drug transporters and / or drug metabolism characteristics at gene or protein expression level. Background of the invention [0002] Pluripotent human stem cells are expected to revolutionize the accessibility to various human cell types. The possibility of propagating pluripotent human blastocyst-derived ...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to view more

Application Information

Patent Timeline
no application Login to view more
Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/08C12N5/071
Inventor N·海因斯B·屈佩尔斯-芒瑟J·埃德斯巴戈B·屈佩尔斯－芒瑟
Owner CELLECTIS SA
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Try Eureka
PatSnap group products

Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic.

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap