Isolation, expansion and use of clonogenic endothelial...

C - Chemistry – Metallurgy – 12 – N

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C12N 5/071 (2010.01) C12N 5/073 (2010.01) C12N 5/0789 (2010.01) A61K 35/14 (2006.01) A61K 35/28 (2006.01) A61K 35/44 (2006.01) C12N 1/04 (2006.01)

Patent

CA 2554094

A hierarchy of endothelial colony forming cells (EPCs) was identified from mammalian cord blood, umbilical vein and aorta. A newly isolated cell named high proliferative potential - endothelial colony forming cell (HPP-ECFC) was isolated and characterized. Single cell assays were developed that test the proliferative and clonogenic potential of endothelial cells derived from cord blood, or from HUVECs and HAECs. EPCs were found to reside in vessel walls. Use of a feeder layer of cells derived from high proliferative potential- endothelial colony forming cells (HPP-ECPCS) from human umbilical cord blood, stimulates growth and survival of repopulating hematopoietic stem and progenitor cells. Stimulation of growth and survival was determined by increased numbers of progenitor cells in in vitro cultures and increased levels of human cell engraftment in the NOD/SCIID immunodeficient mouse transplant system.

L'invention concerne des cellules endothéliales formant des colonies (progéniteurs de cellules endothéliales) identifiées dans le sang prélevé dans cordon ombilical, dans la veine ombilicale, et dans l'aorte d'un mammifère. L'invention concerne également une cellule nouvellement isolée et caractérisée, appelée HPP-ECFC (high proliferative potential - endothelial colony forming cell : cellule endothéliale formatrice de colonie à pouvoir de prolifération élevé), ainsi que des tests portant sur une seule cellule, permettant de mesurer le pouvoir prolifératif et clonogène des cellules endothéliales humaines dérivées du sang du cordon ombilical, de la veine ombilicale ou de l'artère. On a découvert que ces progéniteurs de cellules endothéliales résidaient dans les parois de vaisseaux sanguins. L'invention porte en outre sur l'utilisation d'une couche nutritive (feeder layer) de cellules dérivées des cellules HPP-ECPCS provenant du cordon ombilical humain, laquelle stimule la croissance et la survie des cellules souches et des progéniteurs de repeuplement. La stimulation de la croissance et de la survie a été mesurée à partir de l'accroissement du nombre de progéniteurs dans les cultures in vitro, et de l'accroissement des taux de prises de greffe de cellules humaines dans le système de transplantation faisant appel à la souris immunodéficiente NOD/SCID.

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