Capsules containing cells with hematopoietic potential

Patent metadata

Field	Value
Publication number	US-2016333310-A1
Application number	US-201415103655-A
Country	US
Kind code	A1
Filing date	Dec 12, 2014
Priority date	Dec 13, 2013
Publication date	Nov 17, 2016
Grant date	—

Abstract

The present invention relates to a capsule comprising at least one cell with hematopoietic potential, said capsule being formed with a liquid core and at least one gelled shell encapsulating totally the liquid core at its periphery, to the use of such a capsule for producing ex vivo enucleated erythroid cells as well as an ex vivo method for producing enucleated erythroid cells using said capsule.

First claim

1 . A capsule comprising a liquid core, a gelled shell totally encapsulating the liquid core at its periphery, the gelled shell being able to retain the liquid core when the capsule is immersed in a gas, the gelled shell comprising at least one gelled polyelectrolyte and at least one surfactant, characterized in that the liquid core comprises at least one cell with hematopoietic potential.

2 . The capsule according to claim 1 , characterized in that it is obtained by applying a method comprising the following steps:
a) separately conveying in a jacket a first physiologically acceptable liquid solution containing at least one cell with hematopoietic potential and of a second liquid solution containing a liquid polyelectrolyte able to be gelled; b) forming, at the outlet of the jacket, a series of drops, each drop comprising a central core formed with said first solution and a peripheral film formed with said second solution and totally covering the central core; c) immersing each drop into a gelling solution containing a reagent able to react with the polyelectrolyte of the film so as to have it pass from a liquid state to a gelled state and forming the gelled shell, the central core forming the liquid core; d) recovering the formed capsules; the second solution containing at least one surfactant before its contact with the first solution.

3 . The capsule according to claim 2 , characterized in that the ratio of the flow rate of the first solution to the flow rate of the second solution at the outlet of the jacket is comprised between 1 and 200, advantageously between 10 and 200, the gelled shell having a thickness comprised between 0.1% and 10%, advantageously between 0.1% and 2% of the diameter of the capsule, after recovering the formed capsules.

4 . The capsule according to claim 2 , characterized in that the drops formed by co-extrusion in the jacket fall by gravity through a volume of air in the gelling solution.

5 . The capsule according to claim 2 , characterized in that the first physiologically acceptable liquid solution comprises a saline solution, a buffer solution, a physiologically acceptable viscosifying solution, a physiologically acceptable excipient, advantageously a thickener or a rheology modifier, and/or some culture medium.

6 . The capsule according to claim 1 , characterized in that it further comprises an intermediate shell totally encapsulating at its periphery the liquid core, said intermediate shell being itself encapsulated totally at its periphery by the gelled shell.

7 . The capsule according to any of claim 1 , characterized in that said at least one cell with hematopoietic potential is a hematopoietic stem cell and/or an erythroid progenitor cell and/or an erythroid precursor.

8 . The capsule according to any of claim 1 , characterized in that said at least one cell with hematopoietic potential or hematopoietic stem cell or erythroid progenitor cell or erythroid precursor is a human cell or precursor.

9 . (canceled)

10 . (canceled)

11 . An ex vivo method for producing enucleated erythroid cells comprising the culture of cells with hematopoietic potential contained in at least one capsule as defined in claim 1 , under conditions allowing the production of enucleated erythroid cells.

12 . The method according to claim 9 , characterized in that the cells with hematopoietic potential are cultivated in a culture medium comprising:
e) insulin at a concentration comprised between 1 and 50 μg/ml; f) transferrin at a concentration comprised between 100 and 2,000 μg/ml; and g) plasma or serum at a concentration comprised between 1 and 30%.

13 . The method according to claim 10 , characterized in that the culture medium also comprises EPO and/or SCF and/or IL-3 and/or hydrocortisone.

14 . The method according to claim 10 , characterized in that the culture medium also comprises at least one of the following compounds: TPO, FLT3, BMP4, VEGF-A 165 and IL-6.

15 . The method according to claim 10 , characterized in that the culture medium also comprises Iscove Dulbecco modified medium, completed with glutamine or a peptide containing glutamine.

16 . The method according to claim 9 , characterized in that the enucleated erythroid cells are reticulocytes and/or erythrocytes.

17 . The method according to claim 9 , characterized in that the cells with hematopoietic potential are hematopoietic stem cells and/or erythroid progenitor cells and/or erythroid precursors.

Assignees

• Etablissement Français Du Sang
• Univ Pierre Etmarie Curie (Paris 6)
• Ecole Supérieure De Physique Et De Chimie Ind De La Ville De Paris
• Hopitaux Paris Assist Publique
• Centre Nat De La Rech Scient (C N R S )

Inventors

• Giarratana Maria Catalina
• Douay Luc
• Domejean Hugo
• Bremond Nicolas
• Brouchon Julie Victoire
• Baudry Jean
• Bibette Jérôme

Classifications

• C12N2501/2303

  Interleukin-3 (IL-3) · CPC title

• C12N2501/91

  Heparin · CPC title

• C12N5/0641

  Erythrocytes · CPC title

• C12N5/0012 Primary

  Cell encapsulation · CPC title

• C12N2501/14

  Erythropoietin [EPO] · CPC title