Skip to main content

Rat Mesenchymal Stem Cell Functional Identification Kit

Key Product Details

Verification of Multipotency using the Rat Mesenchymal Stem Cell Functional Identification Kit.
Catalog #
Availability
Size / Price
Qty
Loading...
SC020

Assay Procedure

Refer to the product datasheet for complete product details.

Briefly, rat MSC multipotency is verified using the following in vitro differentiation procedure:

  • Culture multipotent cells of interest
  • Induce adipocyte, chondrocyte, and osteocyte differentiation using media supplements
  • Evaluate differentiation using mature phenotype marker antibodies and fluorescent ICC
 

 

Reagents Provided

Reagents supplied in the Rat Mesenchymal Stem Cell Functional Identification Kit (Catalog # SC020):

  • Adipogenic Supplement
  • Chondrogenic Supplement
  • Osteogenic Supplement
  • ITS Supplement
  • Adipocyte marker: Goat Anti-Mouse FABP4 Antigen-affinity Purified Polyclonal Antibody
  • Chondrocyte marker: Goat Anti-Human Aggrecan Antigen-affinity Purified Polyclonal Antibody
  • Osteocyte marker: Mouse Anti-Human Osteocalcin Monoclonal Antibody

Note: The quantity of each media supplement in this kit is sufficient to make 50 mL of media for differentiation. 50 mL can be used for 16 wells of a 24-well plate for osteogenic and adipogenic lineages and 10 chondrocyte pellets.

 

Other Supplies Required

Reagents

  • StemXVivo® Osteogenic/Adipogenic Base Media (Catalog # CCM007 or equivalent)
  • D-MEM/F-12 (1X)
  • Phosphate Buffered Saline (PBS)
  • Penicillin-Streptomycin-Glutamate (100X)
  • 4% Paraformaldehyde in PBS
  • 1% BSA in PBS
  • Mounting medium (Catalog # CTS011 or equivalent)
  • NorthernLights 557-conjugated Donkey Anti-Goat IgG Secondary Antibody (Catalog # NL001 and NL007 or equivalent)
  • 0.3% Triton® X-100, 1% BSA, 10% normal donkey serum in PBS
  • 1% BSA, 10% normal donkey serum in PBS
  • Fibronectin (optional; Human Fibronectin, Catalog # 1918-FN, Bovine Fibronectin, (Catalog # 1030-FN, or equivalent)
  • Deionized or distilled water

Materials

  • Rat MSCs
  • 24-well culture plates
  • 12 mm coverslips (Carolina Biologicals, Catalog # 633009 or equivalent)
  • 15 mL centrifuge tubes
  • Pipettes and pipette tips
  • Serological pipettes
  • Glass slides
  • Fine pointed curved forceps
  • Liquid barrier pen

Equipment

  • 37 °C and 5% CO2 incubator
  • Centrifuge
  • Hemocytometer
  • Inverted microscope
  • 2 °C to 8 °C refrigerator
  • 37 °C water bath
  • Fluorescence microscope
  • Cryostat
 

 

Procedure Overview

This protocol has been tested using bone marrow- and/or adipose tissue-derived MSCs. If using a different tissue source or cell line, the protocol below may need to be optimized.

Adipogenic Differentiation

Plate 2.1 x 104 MSCs/cm2 in StemXVivo® Osteogenic/Adipogenic Base Media.

Culture cells to 100% confluency.

Culture cells to 100% confluency.

Replace the medium with Adipogenic Differentiation Medium to induce adipogenesis.

Replace the medium with Adipogenic Differentiation Medium to induce adipogenesis.

Every 3-4 days, replace with fresh Adipogenic Differentiation Medium.

After 7-21 days, adipocytes can be fixed.

 

ICC detection of FABP4.

ICC detection of FABP4.

Osteogenic Differentiation

 

Plate 4.2 x 103 MSCs/cm2 in StemXVivo® Osteogenic/Adipogenic Base Media.

Culture cells to 50-70% confluency.

Plate 4.2 x 103 MSCs/cm 2 in StemXVivo Osteogenic/Adipogenic Base Media.

Replace the medium with Osteogenic Differentiation Media to induce osteogenesis.

Replace the medium with Osteogenic Differentiation Medium to induce osteogenesis.

Every 3-4 days, replace with fresh Osteogenic Differentiation Medium.

After 14-21 days, osteocytes can be fixed.

 

ICC detection of Osteocalcin.

ICC detection of Osteocalcin.

Chondrogenic Differentiation

 

Transfer 2.5 x 104 MSCs to a 15 mL conical tube.

Centrifuge and resuspend the cells in Chondrogenic Differentiation Media.

Centrifuge the cells but do not remove the medium.

Transfer 2.5 x 104 MSCs to a 15 mL conical tube.

Every 2-3 days, replace with fresh Chondrogenic Differentiation Media.

After 14-21 days, the chondrogenic pellet can be fixed.

Every 2-3 days, replace with fresh Chondrogenic Differentiation Media.

Cryosection the chondrogenic pellet.

 

ICC detection of Aggrecan.

ICC detection of Aggrecan.

Citations (2)

Showing  1 - 2 of 2 citations Showing All
Filter By:
Search Title:

Citations are publications that use Bio-Techne products. Selected citations for Rat Mesenchymal Stem Cell Functional Identification Kit include:

There are no citations that match your criteria.

Showing  1 - 2 of 2 citations Showing All

Product Documents

Certificate of Analysis

To download a Certificate of Analysis, please enter a lot number in the search box below.

Note: Certificate of Analysis not available for kit components.

Complete Your Research

Complete Your Experiment