Pancreatic Cell Differentiation Medium

This three-dimensional (3-D) semi-solid media contains the necessary components for the differentiation of dissociated single pancreatic cells into insulin-producing cells from murine adult and postnatal samples.

Highlights:

  • Semi-solid medium format allows uniform presentation of extracellular matrix components and growth factors to cells, enabling progenitors to proliferate and differentiate in vitro
  • Allows for easy detection and quantitative analysis of functional progenitors, within a heterogeneous population of cells
  • Semi-solid media are spread evenly across the culture well, permitting easy counting of colonies or organoids developed from single progenitor cells
  • This medium does not support human cell differentiation
  • This medium does not contain exogenous R-Spondin 1

From the laboratory of H. Teresa Ku, PhD, City of Hope.

The Investigator's Annexe Part of The Investigator's Annexe program.

Catalog Number Product Size AVAILABILITY Price Qty
ECH101
Pancreatic Cell Differentiation Medium, 2.5mL
4 wells In stock
Price: $226.00
ECH102
Pancreatic Cell Differentiation Medium, 12.5mL
20 wells In stock
Price: $847.00
Specifications
Product Type: Buffer or Chemical
Growth Conditions: Pancreatic Cell Differentiation Medium
Formulation: Liquid/Semi-Solid
Phenol Red Indicator: Present
Serum Level: 0.05
Components: Soluble and conditioned media with growth factors and viscous materials
Storage: 4C, do not freeze. Use within 2 weeks of receiving
Shipped: Cold packs
Data

Representative photomicrograph of insulin-expressing "ring" colonies in semi solid culture

Colonies appear as circles when viewed under a phase-contrast microscope, but are in fact cystic in 3D space. Individual colonies can later be assayed for gene and protein expression by RT-PCR and immunohistochemistry analyses.

Provider
From the laboratory of H. Teresa Ku, PhD, City of Hope.
Comments

IMPORTANT: Be sure to use within 2 weeks of receiving material

For detailed information on plating of cells and counting of the resulting colonies, please refer to Winkler M., et al. J Vis Exp. 2011 Nov 28;(57):e3148.

For information on dissociation of adult murine pancreas, please see Jin et al. Proc Natl Acad Sci U S A 110:3907-3912 (2013).

Recommended Protocol:

  1. Important: keep cold all time for the tube(s), cells, and culture plates prior to incubation.
  2. To collect the semi-solid medium, spin the tube provided at 4oC, 300 g for 5 min.
  3. Add cells to the semi-solid medium. For a 4-well experiment-Add 217.5 uL cells (in DMEM/F12 with 5% fetal calf serum) to the semi-solid medium. (Note, the settled amount is not sufficient for 5 wells). For a 20-well experiment-Add 1.0875 mL cells to the semi-solid medium. (Leave outside wells with 2 mL of sterile water to keep plate hydrated).
  4. Vigorously shake the tube(s) to mix the contents, followed by rest the tube(s) on ice for 5 min (to allow the bubbles to settle).
  5. Use a 16 gauge needle on a 1 mL syringe to dispense 500 uL to each well. Be sure to use low attachment 24-well plates only. Add the media to the center of the well and it will spread out evenly into the wells.
  6. Incubate cells at 37C and 5% CO2.

NOTE: Up to 25,000 cells can be plated per well in a 24-well dish. However, optimal plating density of cells will vary based of cell populations and should be tested by individual laboratories.

References
  1. Jin L, Feng T, Shih, HP, Zerda R, Luo A, et al. Colony-forming cells in the adult mouse pancreas are expandable in Matrigel and form endocrine/acinar colonies in laminin hydrogel. Proc Natl Acad Sci U S A 110:3907-3912 (2013).
  2. Winkler M, Trieu N, Feng T, Jin L, Walker S, Singh L, Ku HT. A quantitative assay for insulin-expressing colony-forming progenitors. J Vis Exp. 2011 Nov 28;(57):e3148.

If you publish research with this product, please let us know so we can cite your paper.

 
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