US 11,834,673 B2
Method for generating human multipotent mammary stem cells from normal primary breast luminal cells
Abdelilah Aboussekhra, Riyadh (SA); Huda H. Al-Khalaf, Riyadh (SA); and Hazem Ghebeh, Riyadh (SA)
Assigned to KING FAISAL SPECIALIST HOSPITAL & RESEARCH CENTRE, Riyadh (SA)
Filed by 03;KING FAISAL SPECIALIST HOSPITAL & RESEARCH CENTRE, Riyadh (SA)
Filed on Aug. 10, 2021, as Appl. No. 17/398,328.
Application 16/029,858 is a division of application No. 15/461,043, filed on Mar. 16, 2017, granted, now 10,100,275, issued on Oct. 16, 2018.
Application 17/398,328 is a continuation of application No. 16/029,858, filed on Jul. 9, 2018, granted, now 11,512,286.
Prior Publication US 2021/0371819 A1, Dec. 2, 2021
This patent is subject to a terminal disclaimer.
Int. Cl. C12N 5/071 (2010.01); A61K 35/55 (2015.01)
CPC C12N 5/0631 (2013.01) [A61K 35/55 (2013.01); C12N 2501/2308 (2013.01); C12N 2513/00 (2013.01)] 14 Claims
 
1. A method of tissue engineering, transplantation, tissue reconstruction, cell therapy procedure, or treatment of the effects of mastectomy, comprising the steps:
A) generating multipotent mammary stem cells by:
a) isolating and growing primary breast luminal cells in-vitro;
b) dedifferentiating the primary breast luminal cells by treating the primary breast luminal cells with either a conditioned medium from active breast stromal fibroblasts or with interleukin-8 (IL-8) in serum-free medium;
c) continuing the growth of the treated cells until the formation of cellular aggregates;
d) transferring the cells to a stem cell growth medium and growing them further until the formation of mammospheres; and
e) isolating said multipotent mammary stem cells; and
B) applying/administering said multipotent mammary stem cells to a patient in need thereof at a place of the patient's body where a process of tissue engineering, transplantation, tissue reconstruction or cell therapy procedure is desired.