Methods for establishing and improving the survival of a population of pancreatic progenitor or stem cells
In some embodiments, methods for improving the survival of pancreatic β-cell progenitors in culture are provided. Such methods may include contacting a population of pancreatic progenitor cells with an amino acid (aa) sequence comprising IKVAV (SEQ ID NO:1). In other embodiments, methods for (i) verifying the establishment of a population of pancreatic progenitor or stem cells and (ii) methods for generating or establishing a population of pancreatic endocrine progenitor cells in vitro are provided.
1. A method of improving the survival of pancreatic β-cell progenitors in culture comprising:
contacting a population of pancreatic progenitor cells with an artificial extracellular matrix (aECM) protein, the aECM protein comprising
a) a first amino acid sequence comprising a functional portion of elastin comprising [(VPGIG) 2 VPGKG(VPGIG) 2 ] 4 (SEQ ID NO: 3); and
b) a second amino acid sequence comprising SEQ ID NO:2;
wherein the aECM protein improves survival of the population of pancreatic β-cell progenitors, and
wherein the population of pancreatic progenitor cells express one or more pancreatic progenitor cell markers selected from Pdx1, Ngn3, Nkx6.1, Pax4, MafB, c-Met, DCAMKL1, and DCAMKL2.
2. The method of claim 1 , wherein the population of pancreatic progenitor cells is derived from a population of adult pancreatic ductal cells.
3. The method of claim 1 , wherein the aECM protein further comprises a T7 tag.
4. A method of improving the survival of pancreatic β-cell progenitors in culture comprising:
contacting a population of pancreatic progenitor cells with an artificial extracellular matrix (aECM) protein,
wherein the aECM protein comprises MASMTGGQQMG-HHHHHH-MKL{X-Y-VP} 3 LE (SEQ ID NO:4),
wherein X is substituted with a second amino acid sequence comprising a functional portion of the α-chain of laminin comprising IKVAV (SEQ ID NO: 1) and Y is substituted with a first amino acid sequence comprising a functional portion of elastin comprising [(VPGIG) 2 VPGKG(VPGIG) 2 ] 4 (SEQ ID NO: 3),
wherein the aECM protein improves survival of the population of pancreatic β-cell progenitors, and
wherein the population of pancreatic progenitor cells express one or more pancreatic progenitor cell markers selected from Pdx1, Ngn3, Nkx6.1, Pax4, MafB, c-Met, DCAMKL1, and DCAMKL2.
5. The method of claim 4 , wherein the second amino acid sequence is SEQ ID NO:2.
6. The method of claim 4 , wherein the first amino acid sequence is SEQ ID NO:3 and the second amino acid sequence is SEQ ID NO:2.
7. A method of improving the survival of pancreatic β-cell progenitors in culture comprising:
contacting a population of pancreatic progenitor cells with an artificial extracellular matrix (aECM) protein, the aECM protein comprising
a) a first amino acid sequence comprising SEQ ID NO:3; and
b) a second amino acid sequence comprising SEQ ID NO:2;
wherein the aECM protein improves survival of the population of pancreatic β-cell progenitors.
8. The method of claim 7 , wherein the aECM protein further comprises a T7 tag.
9. A method of improving the survival of pancreatic β-cell progenitors in culture comprising contacting a population of pancreatic progenitor cells with an artificial extracellular matrix (aECM) protein comprising MASMTGGQQMG-HHHHHH-MKL{X-Y-VP} 3 LE (SEQ ID NO:4), wherein X is substituted with a second amino acid sequence comprising SEQ ID NO: 1 and Y is substituted with a first amino acid sequence comprising SEQ ID NO: 3, and wherein the aECM protein improves survival of the population of pancreatic β-cell progenitors.
10. A method of improving the survival of pancreatic β-cell progenitors in culture comprising:
contacting a population of pancreatic progenitor cells with an artificial extracellular matrix (aECM) protein, the aECM protein comprising MASMTGGQQMG-HHHHHH-MKL{X-Y-VP} 3 LE (SEQ ID NO:4), wherein
X is a laminin component comprising IKVAV (SEQ ID NO: 1) and Y is an elastin component comprising [(VPGIG) 2 VPGKG(VPGIG) 2 ] 4 (SEQ ID NO: 3), or
X is an elastin component comprising [(VPGIG) 2 VPGKG(VPGIG) 2 ] 4 (SEQ ID NO: 3) and Y is a laminin component comprising IKVAV (SEQ ID NO: 1), and
wherein the aECM protein improves survival of the population of pancreatic β-cell progenitors.
11. A method of improving the survival of pancreatic β-cell progenitors in culture comprising:
contacting a population of pancreatic progenitor cells with an artificial extracellular matrix (aECM) protein, the aECM protein comprising MASMTGGQQMG-HHHHHH-MKL{X-Y-VP} 3 LE (SEQ ID NO:4), wherein:
X comprises LDASMSVQAVDRSANKARIVSKAASA (SEQ ID NO:6) or VPLDYAVTGRGDSPASSKIPIA (SEQ ID NO:7); and
Y is substituted with [(VPGIG) 2 VPGKG(VPGIG) 2 ] 4 (SEQ ID NO: 3);
wherein the aECM protein improves survival of the population of pancreatic β-cell progenitors.