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Improved preparations of adult liver progenitor cells

A technology of liver progenitor cells and cells, applied in the field of improved adult liver progenitor cell preparations, can solve the problem of impossible to determine liver progenitor cells and the like

Inactive Publication Date: 2018-11-09
UNIVERSITE CATHOLIQUE DE LOUVAIN
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, it has not been possible to determine which of the hepatic progenitor cells identified so far are those more suitable for the treatment or use of a given disease, mainly due to the Methods of Effect and Variability in Subsequent Drug Use

Method used

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  • Improved preparations of adult liver progenitor cells
  • Improved preparations of adult liver progenitor cells
  • Improved preparations of adult liver progenitor cells

Examples

Experimental program
Comparison scheme
Effect test

preparation example Construction

[0057] (b) a preparation of primary liver cells producing (a);

[0058] (c) culturing the cells contained in the preparation of (b) onto a support which allows the cells to adhere and grow thereon and the emergence of cell populations;

[0059] (d) passage the cells of (c) at least once; and

[0060] (e) Isolating the population of cells obtained after the passaging of (d) that is positive for the markers specified in the Summary of the Invention.

[0061] With respect to step (a) of the method, the dissociating step comprises obtaining an adult liver or a portion thereof containing an amount of primary cells useful for making HHALPCs together with fully differentiated hepatocytes. Liver primary cells are preferentially isolated from human liver tissue obtained from adult livers.

[0062] The term "liver" refers to the liver organ. The term "part of the liver" generally refers to a tissue sample derived from any part of the liver organ without any limitation on the amount o...

Embodiment 1

[0168] Example 1: Analysis of cell surface proteins on HHALPC

[0169] Materials and Methods

[0170] Isolation and Expansion of HHALPC in Cell Culture

[0171] HHALPC were recovered after primary culture of the hepatic parenchymal cell fraction was achieved after two steps of collagenase perfusion, filtration and low speed centrifugation using five different human donors as described elsewhere (Najimi M et al., 2007). These liver cells are suspended in cryopreservation medium prepared with ViaSpan solution and then maintained in liquid nitrogen by using appropriate vials, bags, or other systems for long-term storage and preservation of human cells. Cryopreserved hepatocyte suspensions were quickly thawed at 37°C and supplemented with 2.5g / L glucose, 0.084g / L bicarbonate and 5000IE / UI / ml heparin Wash twice with 10x volume of 5% human albumin. After centrifugation at 224 g for 10 min at 4 °C, the cell pellet was suspended in the desired cell culture medium.

[0172] HH...

Embodiment 2

[0196] Example 2: Validation of the therapeutic properties of HHALPC

[0197] Materials and Methods

[0198] HHALPC preparation and administration to patients affected by urea cycle disorders

[0199] HHALPC were generated from healthy human liver cell suspension and expanded for 5 passages as shown in Example 1, then harvested, cryopreserved in CryoStor-10 (10% DMSO), and stored in liquid nitrogen. Before use, HHALPC were thawed and washed with albumin solution, then formulated in a 50ml plastic bag in a sterile environment in a GMP facility containing 250x10 in 0.084 sodium bicarbonate, 5% human albumin and 500IU heparin 6 cell suspension of cells. HHALPC was infused intravenously through a percutaneous transhepatic portal catheter at a flow rate of 0.5 to 2 ml / min, under general anesthesia, under radiological and ultrasound guidance, via direct transhepatic puncture of the right / left portal vein to the main artery at the confluence of the splenic and portal veins. Po...

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Abstract

Preparations of adult liver progenitor cells (called HHALPCs) have been manufactured from different human donors and characterized by using cell surface markers that allow identifying HHALPCs preparations and / or the methods for producing them that are most suitable for cell therapy, in particular for treating liver diseases or inherited blood coagulation disorders.

Description

technical field [0001] The present invention relates to adult liver progenitor cells generated using primary liver cells and their use for the medical management of liver disease, inherited blood coagulation disorders or for screening compounds with medical interest. Background technique [0002] The liver is a key organ regulating body homeostasis and is the site of many important metabolic pathways. Damage to just one protein in a complex metabolic pathway can be very harmful. The high presence of important liver enzymes significantly increases the risk of several liver diseases. Current treatments and long-term management are not effective enough. Orthotopic liver transplantation (OLT) is highly invasive, irreversible, limited by a shortage of donor grafts and requires state-of-art surgery. Liver cell transplantation (LCT) is only likely to be effective in the short to medium term due to the quality of the hepatocyte preparation. Further improvements in tolerance to c...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/071A61K35/407
CPCC12N5/067G01N33/5067C12N2502/14C12N2531/00G01N33/5008A61K35/407A61P7/02A61P7/04A61P1/16
Inventor 卡特琳·隆巴尔德皮埃尔-爱德华·多莱艾蒂安·索卡尔穆斯塔法·纳吉米
Owner UNIVERSITE CATHOLIQUE DE LOUVAIN
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