EPHA4-positive human adult pancreatic endocrine progenitor cells

• US 8,377,689 B2
• Filed: 09/24/2010
• Issued: 02/19/2013
• Est. Priority Date: 09/27/2005
• Status: Expired due to Fees

• Current Assignee: ReNeuron Limited (ReNeuron Group Plc)
• Original Assignee: ReNeuron Limited (ReNeuron Group Plc)
• Inventors: Tsang, Wen-Ghih, Wang, Yanping, Tang, Jinghua
• Primary Examiner(s): Kim, Taeyoon
• Application Number:
  US12/890,319
• Publication Number:
  US 20110008887A1
• Time in Patent Office: 879 Days
• Field of Search: 435/325, 435/41, 435/70.1, 435/370, 435/374, 424/93.7
• US Class Current: 435/325
• CPC Class Codes: C12N 2501/117 Keratinocyte growth factors (KGF-1, i.e. FGF-7; KGF-2, i.e. FGF-12)
• CPC Class Codes: C12N 2501/12 Hepatocyte growth factor [HGF]
• CPC Class Codes: C12N 2533/54 Collagen; Gelatin
• CPC Class Codes: C12N 5/0676 Pancreatic cells

Abstract

The invention relates to the discovery of a selective cell surface marker that permits the selection of a unique subset of pancreatic stem cells having a high propensity to differentiate into insulin-producing cells or into insulin-producing cell aggregates.

Claims

1. A cell culture of propagating pancreatic cells produced by the steps of:

(a) isolating pancreatic cells from a pancreas,

(b) a EphA4 cell selection, and

(c) a CD56 cell selection,

wherein the EphA4 cell selection comprises

(1) contacting the pancreatic cells with an EphA4 binding reagent;

(2) selecting pancreatic cells that specifically bind to the EphA4 binding reagent; and

(3) separating the selected pancreatic cells from pancreatic cells that do not bind the EphA4 binding reagent to obtain a culture of propagating EphAr pancreatic cells, and

wherein the CD56 cell selection comprises

(x) contacting the pancreatic cells with a CD56 binding reagent;

(y) selecting pancreatic cells that specifically bind to the CD56 binding reagent; and

(z) separating the selected pancreatic cells from pancreatic cells that do not bind the CD56 binding reagent to obtain a culture of propagating CD56⁺ pancreatic cells,

wherein the propagating pancreatic cells have a capacity to differentiate into insulin producing cells and wherein the CD56 cell selection is performed before or after contacting the pancreatic cells with the EphA4 binding reagent.

2. The cell culture of claim 1, wherein the EphA4 binding reagent is labeled.

3. The cell culture of claim 1, wherein the step of selecting is done by fluorescence activated cell sorting.

4. The cell culture of claim 1, wherein the EphA4 cell selecting is done by panning.

5. The cell culture of claim 1, wherein the EphA4 binding reagent is an antibody that specifically binds to the EphA4 protein.

6. The cell culture of claim 1, wherein the pancreas is from a human.

7. A cell culture of claim 1, wherein the CD56 cell selection is performed before contacting the pancreatic cells with the EphA4 binding reagent.

8. A cell culture of claim 1, wherein the CD56 cell selection is performed after cells binding the EphA4 binding reagent are separated from pancreatic cells that do not bind the EphA4 binding reagent.

9. A cell culture comprising a substantially homogenous preparation of propagating pancreatic cells expressing both CD56 and EphA4, wherein the propagating pancreatic cells have the capacity to differentiate into insulin producing cells.