Transcriptional profiling of stem cells and their multilineage differentiation

• US 7,883,847 B2
• Filed: 03/31/2006
• Issued: 02/08/2011
• Est. Priority Date: 04/01/2005
• Status: Active Grant

• Current Assignee: Wake Forest University Health Sciences
• Original Assignee: Wake Forest University Health Sciences
• Inventors: Soker, Shay, Hipp, Jason
• Primary Examiner(s): LI, QIAN JANICE
• Application Number:
  US11/394,766
• Publication Number:
  US 20060246488A1
• Time in Patent Office: 1,775 Days
• Field of Search: None
• US Class Current: 435/6.16
• CPC Class Codes: C12Q 1/6881 for tissue or cell typing, e.g. human leukocyte antigen [HLA] probes
• CPC Class Codes: C12Q 2600/158 Expression markers
• CPC Class Codes: C12Q 2600/16 Primer sets for multiplex assays

Abstract

The present invention concerns methods of screening cells for differentiation or de-differentiation, and/or for status as a pluripotent or multipotent (e.g., “stem”) cell, by detecting the differential expression (e.g., upregulation, downregulation) of genes.

Claims

1. A method of screening a human amniotic fluid stem cell for differentiation into a hepatogenic-specific cell line, comprising:

(a) providing a human amniotic fluid stem cell for which differentiation is to be determined, then

(b) subjecting said human amniotic fluid stem cell to differentiating conditions; and

then

(c) detecting in said human amniotic fluid stem cell differential expression of at least one hepatogenic specific gene selected from the group consisting of stearoyl CoA desaturase (SCD), and 3-hydroxy-3-methylglutaryl-Coenzyme A reductase (HMGCR),

wherein upregulation of expression of said at least one hepatogenic specific gene indicates differentiation of said human amniotic fluid stem cell into a hepatogenic specific cell line.

2. The method of claim 1, wherein said detecting step comprises detecting upregulation of expression of at least two hepatogenic specific genes selected from the group consisting of SCD and HMGCR;

wherein upregulation of said at least two hepatogenic specific genes indicates differentiation of said human amniotic fluid stem cell into a hepatogenic specific cell line.

3. The method of claim 1, wherein does said amniotic fluid stem cell does not form a teratoma when grown in vivo.

4. The method of claim 1, wherein said amniotic fluid stem cell does not form a teratoma within one month after intraarterial injection into a 6-8 week old mouse at a dose of 5×

10⁶ cells per mouse.

5. The method of claim 1, wherein said amniotic fluid stem cells are isolated from amniotic fluid between 14 and 18 weeks of gestation.

6. The method of claim 1, wherein said detecting step comprises detecting in said human amniotic fluid stem cell differential expression of stearoyl CoA desaturase (SCD).

7. The method of claim 1, wherein said detecting step comprises detecting in said human amniotic fluid stem cell differential expression of 3-hydroxy-3-methylglutaryl-Coenzyme A reductase (HMGCR).