# US 12,215,351 B2 · Methods for inducing hematopoietic stem cell specificity

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- Number: US 12,215,351 B2
- Document ID: US12215351B2
- Status: Granted
- Kind: B2
- Application number: 16/640,403
- Filed: 2018-08-21
- Published: 2020-11-19
- Granted: 2025-02-04
- Earliest claimed: 2017-08-22
- Assignees: Children's Medical Center Corporation
- Applicants: The Children's Medical Center Corporation
- Inventors: Leonard I. Zon; Joseph Mandelbaum
- Web page: https://patentgrep.com/patent/US12215351B2

## CPC

- C12N 5/0647: {Haematopoietic stem cells; Uncommitted or multipotent progenitors}
- A61K 31/404: Indoles, e.g. pindolol
- A61K 31/426: 1,3-Thiazoles
- A61K 31/444: containing a six-membered ring with nitrogen as a ring heteroatom, e.g. amrinone
- A61K 31/5513: 1,4-Benzodiazepines, e.g. diazepam {or clozapine}
- A61K 35/50: Placenta; Placental stem cells; Amniotic fluid; Amnion; Amniotic stem cells
- A61K 35/545: Embryonic stem cells; Pluripotent stem cells; Induced pluripotent stem cells; Uncharacterised stem cells
- C12N 5/0062: {General methods for three-dimensional culture}
- A61K 35/28: Bone marrow; Haematopoietic stem cells; Mesenchymal stem cells of any origin, e.g. adipose-derived stem cells
- C12N 2501/60: Transcription factors
- C12N 2501/999: Small molecules not provided for elsewhere
- C12N: MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA (microbiological testing media C12Q1/00)
- A61K: PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES (devices or methods specially adapted for bringing pharmaceutical products into particular physical or administering forms A61J3/00; chemical aspects of, or use of materials for deodorisation of air, for disinfection or sterilisation, or for bandages, dressings, absorbent pads or surgical articles A61L; soap compositions C11D)

## Abstract

Described herein are methods for inducing HSC specification in a cell. Aspects of the invention relate to contacting a cell with a Runx1-CBFβ inhibitor for a specified period of time, and then removing the inhibitor from the cell. In some embodiments of any of the aspects, HSC specificity is maintained long-term.

## Claims

### Claim 1

A method for inducing hematopoietic stem cell (HSC) specification in a cell, the method comprising;
a) contacting a cell for a period of time with an agent selected from the group consisting of Ro5-3335, SU-5402, sc-221405, and AI-10; and
b) removing said agent after said period of time, thereby inducing HSC specification in the cell,
wherein the cell is selected from the group consisting of an hemogenic endothelial (HE) cell, an embryonic cell, an embryonic stem cell (ESC), a cell from a dissociated embryo, a cell from a dissociated embryo that is at least 9-hours post fertilization, an embryoid bodies, an induced pluripotent stem cell (iPSC), an aorta-gonad-mesonephros (AGM) cell, a placenta stem cell, an adult stem cell, and an amniotic stem cell,
wherein the concentration of the agent is greater than 2 μM.

### Claim 2 (depends on 1)

The method of claim 1, wherein the contacting is in vitro, ex vivo or in vivo.

### Claim 3 (depends on 2)

The method of claim 2, wherein the contacting is in vitro.

### Claim 4 (depends on 1)

The method of claim 1, wherein the period of time is between 1 and 6 days.

### Claim 5 (depends on 1)

The method of claim 1, wherein the cell is a mammalian cell, human cell, or non-human mammalian cell.

### Claim 6 (depends on 1)

The method of claim 1, wherein the cell of (a) is an isolated cell.

### Claim 7 (depends on 1)

The method of claim 1, wherein the cell of (a) is an iPS cell.

### Claim 8 (depends on 1)

The method of claim 1, wherein the cell of (a) is a human cell.

### Claim 9 (depends on 1)

The method of claim 1, wherein HSC specification is maintained long-term.

### Claim 10 (depends on 9)

The method of claim 9, wherein long-term is at least 3 months.

### Claim 11 (depends on 1)

The method of claim 1, wherein the HSC specification-induced cell is transplanted into a recipient.

### Claim 12 (depends on 1)

The method of claim 1, wherein the HSC specification induced cell differentiates into a blood cell.