REGENERATION METHOD USING SOMATIC CELL NUCLEAR TRANSFER (SCNT) CELL AND BLASTOCYST COMPLEMENTATION

• US 20190327945A1
• Filed: 06/14/2017
• Published: 10/31/2019
• Est. Priority Date: 06/14/2016
• Status: Active Application

• Current Assignee: Riddle Institute For Regenerative Medicine
• Original Assignee: Riddle Institute For Regenerative Medicine
• Inventors: Riddle, Jr., Michael C.
• Application Number:
  US16/309,907
• Publication Number:
  US 20190327945A1
• Time in Patent Office: Days
• CPC Class Codes: A01K 2207/15 Humanized animals
• CPC Class Codes: A01K 2217/05 Animals comprising random inserted nucleic acids (transgenic)
• CPC Class Codes: A01K 2227/10 Mammal
• CPC Class Codes: A01K 2227/105 Murine
• CPC Class Codes: A01K 2267/025 Animal producing cells or organs for transplantation
• CPC Class Codes: A01K 67/0271 Chimeric vertebrates, e.g. comprising exogenous cells
• CPC Class Codes: A01K 67/0273 Cloned vertebrates
• CPC Class Codes: A01K 67/0276 Knock-out vertebrates

Abstract

We disclose that a target organ such as kidney can be regenerated by complementing a developmental deficiency leading to a lack of development of the target organ in a non-human first mammal by injecting a somatic cell nuclear transfer cell (SCNT cell) into a developed blastocyst of the non-human first mammal. We also disclose a method for producing a target organ, using an SCNT cell, in a living body of a non-human first mammal having an abnormality associated with a lack of development of the target organ in a development stage, the target organ produced being derived from a second mammal that is an individual different from the non-human first mammal.

Claims

1. A method for producing a target organ in a living body of a non-human first mammal having an abnormality associated with a lack of development of the target organ in a development stage, the target organ produced being derived from a second mammal that is an individual different from the non-human first mammal, the method comprising:

a) preparing a somatic cell nuclear transfer cell (SCNT cell) derived from the second mammal;

b) transplanting the SCNT cell into a blastocyst of the non-human first mammal, to yield a chimeric embryo;

c) developing the chimeric embryo in a uterus of a non-human third mammal to obtain at least one offspring comprising the target organ; and

d) obtaining the target organ from the at least one offspring.

2. The method according to claim 1, wherein the second mammal is a human, a rat, or a mouse.

3. The method according to claim 1, wherein the second mammal is a human.

4. The method according to claim 1, wherein the organ to be produced is selected from a pancreas, a kidney, a thymus, and a hair.

5. The method according to claim 1, wherein the non-human first mammal is a mouse, a rat, a monkey, or a pig.

6. The method according to claim 5, wherein the non-human first mammal is a mouse, and the mouse is one or more of a Sall 1 knockout mouse, a Pdx1-Hes1 transgenic mouse, a Pdx1 knockout mouse, and a nude mouse.

7. The method according to claim 1, wherein the blastocyst is prepared from an oocyte of the non-human first mammal in which one or more genes associated with the abnormality have been silenced by a CRISPR technique and a sperm of the non-human first mammal in which one or more genes associated with the abnormality have been silenced by a CRISPR technique.

8. The method according to claim 1, wherein the target organ is completely derived from the second mammal.

9. The method according to claim 1, wherein the second mammal and the non-human first mammal are in a xenogeneic relationship.

10. A non-human first mammal having an abnormality associated with a lack of development of a target organ in a development stage, the mammal being produced by a method comprising:

a) preparing an SCNT cell derived from a second mammal that is an individual different from the non-human first mammal;

b) transplanting the SCNT cell into a blastocyst of the non-human first mammal, to yield a chimeric embryo; and

c) developing the chimeric embryo in a uterus of a non-human third mammal to obtain at least one offspring comprising the target organ.

11. A kit for producing a target organ, the kit comprising:

A) a non-human first mammal having an abnormality associated with a lack of development of the target organ in a development stage; and

B) an SCNT cell derived from a second mammal that is an individual different from the non-human first mammal.

12. A method for producing one or more of a target organ and a target body part, the method comprising:

providing a non-human animal which includes a deficiency-responsible gene coding for a factor which causes a deficiency of one or more of an organ and a body part and gives one or more of no possibility of survival and difficulty in survival if the factor functions, and in which the one or more of an organ and a body part is complemented by blastocyst complementation, the deficiency-responsible gene coding for a factor which causes a deficiency of the one or more of a target organ and a target body part;

growing an ovum obtained from the non-human animal into a blastocyst;

introducing a target somatic cell nuclear transfer cell (SCNT cell) into the blastocyst so as to produce a chimeric blastocyst, the target SCNT cell having a desired genome capable of complementing a deficiency caused by the deficiency-responsible gene;

producing an individual from the chimeric blastocyst; and

obtaining the one or more of a target organ and a target body part from the individual.

13. The method according to claim 12, wherein producing the individual further comprises developing the chimeric blastocyst in a uterus of a non-human third mammal to obtain at least one offspring comprising the target organ, and obtaining the target organ from the at least one offspring.

14. The method according to claim 12, wherein the target SCNT cell is derived from a human.

15. The method according to claim 12, wherein the one or more of a target organ and a target body part is selected from a pancreas, a kidney, a thymus, and a hair.

16. The method according to claim 12, wherein the non-human animal is a mouse, a rat, a monkey, or a pig.

17. The method according to claim 16, wherein the non-human animal is a mouse, and the mouse is one or more of a Sall 1 knockout mouse, a Pdx1 knockout mouse, a Pdx1-Hes1 transgenic mouse, and a nude mouse.

18. The method according to claim 12, wherein the one or more of a target organ and a target body part is completely derived from the target SCNT cell.

19. The method according to claim 12, further comprising preparing the SCNT cell, wherein the SCNT cell is derived from a second mammal.

20. The method according to claim 19, wherein the second mammal and the non-human animal are in a xenogeneic relationship.

21. The method according to claim 12, wherein the blastocyst is prepared from an oocyte of the non-human first mammal in which one or more genes, including the deficiency-responsible gene, have been silenced by a CRISPR technique and a sperm of the non-human first mammal in which one or more genes, including the deficiency-responsible gene, have been silenced by a CRISPR technique.

22. A kit for producing one or more of a target organ and a target body part, the kit comprising:

A) a non-human animal which includes a gene coding for a factor which causes a deficiency of one or more of an organ and a body part and gives one or more of no possibility of survival and difficulty in survival if the factor functions, and in which the one or more of an organ and a body part is complemented by complement; and

B) an SCNT cell derived from a second mammal that is an individual different from the non-human animal.

23. The kit according to claim 22, wherein the non-human animal and the second mammal are in a xenogeneic relationship.