ZSCAN4 as a marker for pancreatic stem cells and progenitor cells and use thereof
ZSCAN4, a gene expressed in ES cells and 2-cell stage embryos, has been previously shown to regulate telomere elongation and genome stability in mouse ES cells. It is disclosed herein that in the adult human pancreas, a small number of ZSCAN4-positive cells are present among cells located in the islets of Langerhans, acini, and ducts. These data disclosed herein indicates that expression of ZSCAN4 is a marker for rare stem/progenitor cells in adult human pancreas. Thus, provided herein is a method of isolating pancreatic stem cells or progenitor cell from a sample by detecting expression of ZSCAN4. Also provided is a method of treating diabetes by isolating ZSCAN4 + pancreatic stem cells or progenitor cells, expanding the cells in vitro and transplanted the expanded cells into the subject. The expanded ZSCAN4 + cells can optionally be differentiated into pancreatic β cells before transplanting the cells into the subject. Further provided is an in vivo method of expanding a population of pancreatic stem cells or progenitor cells in a subject by delivering a ZSCAN4 protein, a ZSCAN4 nucleic acid sequence, or an agent that increases expression of ZSCAN4 to the pancreas of the subject.
1. A method of isolating pancreatic stem cells, pancreatic progenitor cells, or pancreatic stem cells and pancreatic progenitor cells that express a zinc finger and SCAN domain containing protein 4 (ZSCAN4 protein) from a sample comprising pancreatic tissue, the method comprising:
(i) detecting expression of the ZSCAN4 protein, an mRNA encoding the ZSCAN4 protein, an SSEA3 protein, an mRNA encoding the SSEA3 protein, a Tcstv1/3 protein, or an mRNA encoding the Tcstv1/3 protein in cells of the sample; and
(ii) isolating the cells that express the ZSCAN4 protein, the mRNA encoding the ZSCAN4 protein, the SSEA3 protein, the mRNA encoding the SSEQA3 protein, the Tcstv1/3 protein or an mRNA encoding the Tcstv1/3 protein, thereby isolating pancreatic stem cells, pancreatic progenitor cells, or pancreatic stem cells and pancreatic progenitor cells from the sample.
2. The method of claim 1 , wherein detecting expression of the ZSCAN4 protein comprises contacting the sample with an antibody specific for the ZSCAN4 protein.
3. The method of claim 1 , wherein detecting expression of the mRNA encoding the ZSCAN4 protein is by PCR.
4. The method of claim 1 , wherein detecting expression of the SSEA3 protein or the Tcstv1/3 protein comprises contacting the sample with an antibody specific for the SSEA3 protein or the Tcstv1/3 protein, respectively.
5. The method of claim 4 , wherein the antibody is specific for the SSEA3 protein.
6. The method of claim 4 , wherein the antibody is specific for the Tcstv1/3 protein.
7. The method of claim 1 , wherein detecting expression of ZSCAN4 comprises transfecting the cells of the sample with a vector comprising a ZSCAN4 promoter operably linked to a reporter gene or a selectable marker.
8. The method of claim 7 , wherein the ZSCAN4 promoter is a mouse Zscan4c promoter.
9. The method of claim 8 , wherein the Zscan4c promoter comprises the nucleic acid sequence set forth as nucleotides 906-4468 of SEQ ID NO: 15.
10. The method of claim 7 , wherein the reporter gene encodes a fluorescent protein.
11. The method of claim 10 , wherein the fluorescent protein is a green fluorescent protein, or a variant thereof.
12. The method of claim 7 , wherein the vector comprises the nucleic acid sequence set forth as SEQ ID NO: 15.
13. The method of claim 7 , wherein the selectable marker is an antibiotic resistance gene.
14. The method of claim 13 , wherein the antibiotic resistance gene is a puromycin-resistance gene.
15. The method of claim 1 , wherein the sample comprises human pancreatic tissue obtained by biopsy.
16. The method of claim 1 , wherein the method further comprises detecting expression of LGR5 or BMI1, or both, in cells of the sample, and isolating cells that also express LGR5 or BMI1, or both.