Isolated populations of adult renal cells and methods of isolating and using same

Patent metadata

Field	Value
Publication number	US-10870834-B2
Application number	US-201816045784-A
Country	US
Kind code	B2
Filing date	Jul 26, 2018
Priority date	May 13, 2010
Publication date	Dec 22, 2020
Grant date	Dec 22, 2020

Abstract

A method of generating a nephrospheroid is disclosed. The method comprises culturing human adult kidney cells in a culture medium under non-adherent conditions. Uses thereof and other renal cell populations are also disclosed.

First claim

We claim:

1. A method for forming tubular structures in the kidney of a subject in need thereof, comprising:
expanding human kidney epithelial cells from a human adult kidney in a first culture medium under adherent conditions, wherein the first culture medium is a serum containing medium comprising at least one growth factor comprising IMDM supplemented with 10% FBS, 1% L-glutamine, 1% Pen-Strep, and growth factors: 50 ng/ml of bFGF, 50 ng/ml of EGF, and 5 ng/ml of SCF;
culturing the cells in a second culture medium under non-adherent conditions, thereby generating a plurality of nephrospheroid cells, wherein the second culture medium is a serum free medium comprising 500 ml DMEM:F12 in a 1:1 ratio, 1% pen-strep, 2 ml B27 supplement, 4 μg/ml heparin, 1% non-essential amino acids, 1% sodium pyruvate, 1% L-glutamine, 1 ml lipid mix, 5 ml of 100× N2 supplement, 10 ng/ml FGF, 20 ng/ml EGF, and 5 ml growth factor mix; wherein a 200 ml solution of growth factor mix comprises 100 ml DMEM:F12, 4 ml of 30% glucose, 200 mg transferin, 50 mg insulin in 20 ml of water, 19.33 mg putrescine in 20 ml ddw, 200 μl of 0.3 mM sodium selenite, and 20 μl of 2 mM progesterone;
isolating a population of nephrospheroid cells from the second culture medium; and
administering to the subject a therapeutically effective amount of the isolated nephrospheroid cells by injection to a damaged kidney of the subject.

2. The method according to claim 1 further comprising removing kidney cells from a subject suffering from renal damage, prior to expanding the cells.

3. The method according to claim 1 further comprising testing a nephrospheroid cell for the presence of a renal progenitor cell marker.

4. The method according to claim 3 wherein the nephrospheroid cells express NCAM.

5. The method according to claim 4 wherein the nephrospheroid cells have increased levels of sall1, pax2, six2 and WT1.

6. The method according to claim 1 wherein the isolated nephrospheroid cells are administered with a pharmaceutically acceptable carrier.

7. The method according to claim 1 wherein the subject suffers from renal failure.

8. The method according to claim 1 wherein the subject suffers from acute or chronic kidney disease, diabetic nephropathy, renal disease associated with hypertension, hypertensive acute tubular injury, interstitial nephritis, Aplasia, dysplasia, obstructive uropathy, reflux nephropathy, Juvenile nephronophtisis, autosomal recessive polycystic kidney disease, Alport, Cystinosis, Primary Hyperoxaluria, Glomerulonephritides, Focal Segmental Glomerulosclerosis, systemic lupus erythematosus, Henoch-Schönlein purpura, or Hemolytic-uremic syndrome.

9. The method according to claim 1 wherein at least 50% of the nephrospheroid cells are NCAM+.

10. The method according to claim 9 wherein the nephrospheroid cells are EpCAM+.

11. The method according to claim 1 wherein the cells from the human adult kidney are expanded for two passages or fewer.

12. The method according to claim 1 wherein the tubular structures are proximal and distal tubules.

Assignees

• Tel Hashomer Medical Res Infrastructure & Services Ltd
• Univ Ramot

Inventors

• Dekel Benjamin
• Harari-Steinberg Orit
• Buzhor Ella

Classifications

• C12N5/0686 Primary

  Kidney cells · CPC title

• C12N2533/40

  Polyhydroxyacids, e.g. polymers of glycolic or lactic acid (PGA, PLA, PLGA); Bioresorbable polymers · CPC title

• C12N2500/90

  Serum-free medium, which may still contain naturally-sourced components · CPC title

• C12N2501/11

  Epidermal growth factor [EGF] · CPC title

• C12N2501/392

  Sexual steroids · CPC title