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.
Part of The Investigator's Annexe program.
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.
Part of The Investigator's Annexe program.
| Catalog Number | Product | DataSheet | Size | AVAILABILITY | Price | Qty |
|---|
| Product Type: | Buffer or Chemical |
| Growth Conditions: | Pancreatic Cell Differentiation Medium |
| Phenol Red Indicator: | Present |
| Serum Level: | 0.05 |
| Format: | Liquid/Semi-Solid |
| 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 |
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.
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:
- Important: keep cold all time for the tube(s), cells, and culture plates prior to incubation.
- To collect the semi-solid medium, spin the tube provided at 4oC, 300 g for 5 min.
- 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).
- 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).
- 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.
- 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.
- 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).
- 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.
