Process for kidney cell manufacture and treatment

Patent metadata

Field	Value
Publication number	US-2023051170-A1
Application number	US-202117795270-A
Country	US
Kind code	A1
Filing date	Jan 25, 2021
Priority date	Jan 28, 2020
Publication date	Feb 16, 2023
Grant date	—

Abstract

Embodiments described herein relate to a method for preparing cultured cells, the method comprising: obtaining kidney tissue from a human subject; mechanically dissociating the tissue; subjecting the tissue to enzymatic digestion; incubating the tissue with media in a cell culture plate to form cultured cells.

First claim

1 . A method for manufacture of cell nephroforms for kidney therapy, the method comprising:
a. obtaining kidney tissue from a human subject; b. mechanically dissociating the tissue; c. subjecting the tissue to enzymatic digestion, wherein the kidney tissue is not filtered or strained after mechanical and enzymatic digestion; d. incubating the tissue with serum containing media (SCM) having at least one growth factor in a cell culture plate to form cultured cells; e. harvesting the cultured cells from the cell culture plate using a dissociating agent; f. reseeding the cultured cells in tissue culture flasks, and incubating the cells under non-adherent conditions, to form nephroforms.

2 . The method according to claim 1 wherein the kidney tissue is a piece of kidney tissue weighing between 2 mg and 100 mg.

3 . The method according to claim 2 wherein the piece of kidney tissue weighs between 2 mg and 50 mg.

4 . The method according to claim 1 wherein the tissue is obtained from the subject using a biopsy.

5 .- 7 . (canceled)

8 . The method according to claim 1 wherein the tissue is subjected to mechanical and enzymatic digestion while maintaining the same weight of the kidney tissue.

9 . (canceled)

10 . The method according to claim 1 wherein the tissue is washed with a buffer solution before incubating the tissue with SCM.

11 .- 25 . (canceled)

26 . A method for preparing cultured cells, the method comprising:
a. obtaining kidney tissue from a human subject; b. mechanically dissociating the tissue; c. subjecting the tissue to enzymatic digestion; and d. incubating the tissue with media in a cell culture plate to form cultured cells, wherein the kidney tissue is not filtered or strained after mechanical and enzymatic digestion.

27 . The method according to claim 26 wherein the kidney tissue is a piece of kidney tissue weighing between 2 mg and 100 mg.

28 . The method according to claim 27 wherein the piece of kidney tissue weighs between 2 mg and 50 mg.

29 . The method according to claim 26 wherein the tissue is obtained from the subject using a biopsy.

30 . The method according to claim 26 wherein the tissue is subjected to mechanical and enzymatic digestion while maintaining the same weight of the kidney tissue.

31 . (canceled)

32 . The method according to claim 26 wherein the tissue is washed with a buffer solution before incubating the tissue with media.

33 . The method according to claim 26 wherein the cell culture plate is a 35 mm to 100 mL culture plate/tube.

34 . The method according to claim 26 wherein the media is SCM, comprising fetal bovine serum.

35 . The method according to claim 34 wherein the SCM further comprises at least one growth factor selected from the group consisting of:
fibroblast growth factor (FGF) stem cell factor (SCF) (5 ng/mL), and epidermal growth factor (EGF).

36 . The method according to claim 26 wherein the kidney tissue is incubated for between 7 and 10 days in step d.

37 . (canceled)

38 . The method according to claim 26 wherein the medium is changed over the course of the incubation.

39 . The method according to claim 38 wherein the medium is changed after 2 days.

40 . The method according to any claim 26 wherein the kidney tissue is incubated until cells achieve confluence.

41 . The method according to claim 26 , further comprising:
e. harvesting the cultured cells from the cell culture plate using a dissociating agent.

42 .- 45 . (canceled)

Assignees

• Kidneycure Ltd
• Tel Hashomer Medical Res Infrastructure & Services Ltd

Inventors

• Simanovsky Masha
• Dekel Benjamin

Classifications

• C12N2533/90

  Substrates of biological origin, e.g. extracellular matrix, decellularised tissue · CPC title

• A61L2430/26

  for kidney reconstruction · CPC title

• A61P13/12

  of the kidneys · CPC title

• A61K9/0019

  Injectable compositions; Intramuscular, intravenous, arterial, subcutaneous administration; Compositions to be administered through the skin in an invasive manner (non-active ingredients are additionally classified in A61K47/00) · CPC title

• C12N2509/00

  Methods for the dissociation of cells, e.g. specific use of enzymes · CPC title