Accelerate research, empower industry

Product Name：Chondrogenic Differentiation Medium for Mesenchymal Stem Cell

Catalog No.：GUMD-D203

Size：200 mL

Product Introduction

Under the action of induction medium, mesenchymal stem cells (MSCs) gradually differentiate into chondrocytes. Chondrocytes are surrounded by an extracellular matrix rich in proteoglycans, which serves as a marker of chondrogenic differentiation and can be stained blue-green with Alcian blue. This product is suitable for chondrogenic induction of MSCs and can significantly improve induction efficiency.

Product Features

· Stable performance, significantly enhancing the chondrogenic induction efficiency of MSCs.

· Suitable for both 2D and 3D chondrogenic induction of MSCs.

· Includes staining solution for convenience.

· Ready-to-use product, saving time and effort.

Example: Chondrogenic Induction Steps (3D Induction)

(The following steps are for reference only. For details, refer to the manual.)

1. Preparation of Mesenchymal Stem Cells:

· Digest cells in the logarithmic growth phase, count them, and transfer 3×10⁵ cells to a 15 mL centrifuge tube. Centrifuge at 250 ×g for 4 min.

· Discard the supernatant, add 0.5 mL of chondrogenic induction differentiation basal medium, resuspend the cells, and centrifuge at 150 ×g for 5 min.

· Carefully discard the supernatant, add 0.5 mL of chondrogenic induction differentiation induction medium, resuspend the cells, and centrifuge at 150 ×g for 5 min.

· Loosen the cap of the 15 mL centrifuge tube slightly and incubate at 37°C with 5% CO₂.

2. Cell Differentiation Induction:

· After 24 hours, observe the aggregation of cell pellets. If significant changes are observed, gently dislodge the cell pellet from the bottom of the tube to ensure it is fully immerse in the induction medium.

· Continue culturing at 37°C with 5% CO₂ for approximately 21 days, replacing the freshly prepared chondrogenic induction differentiation induction medium every 2 days.

· Monitor the formation and surface smoothness of the cell pellet to determine the termination time of induction and proceed with staining identification.

3. Staining Identification:

a) Fixation of Chondrogenic Pellet: Transfer the pellet to an EP tube, wash twice with 1× PBS, and fix in an appropriate amount of 4% neutral formaldehyde solution.

b) Paraffin Embedding and Sectioning: Embed the pellet in paraffin and prepare sections.

c) Alcian Blue Staining: Deparaffinize the sections, stain with Alcian blue for 30 min, rinse under running tap water for 2 min, and wash once with distilled water.

d) Induction Evaluation: Observe the chondrogenic staining effect under a microscope, capture images, and evaluate the induction. Successful induction is indicated by the blue-green staining of acidic mucopolysaccharides in the cartilage tissue with Alcian blue.

NOTE: The chondrogenic differentiation level of mesenchymal stem cells varies depending on cell type, donor source, culture conditions, passage number, cell state, and differentiation time.

Example: Chondrogenic Induction Steps (2D Induction)

(The following steps are for reference only. For details, refer to the manual.)

1.Digest cells in the logarithmic growth phase, count them, and resuspend in chondrogenic induction differentiation medium. Adjust the cell density to 1–2.0×10⁷ cells/mL after centrifugation.

2.Place a 20 µL droplet (containing 4×10⁵ cells) in the center of a 24-well plate. Incubate at 37°C for 3 hours to allow cell attachment.

3.Add 200 µL of induction differentiation medium and culture normally, replacing the medium every 2–3 days. Induce for 21–28 days, observing changes in cell morphology.

4.Cell Fixation: Remove the medium, wash once with 1× PBS, discard, and cover the dish bottom with 4% neutral formaldehyde solution. Fix at room temperature for 30–60 min, discard the fixative, and wash twice with 1× PBS.

5.Alcian Blue Staining: Add an appropriate amount of staining solution to the well, stain in the dark for 30 min, discard the staining solution, wash twice with 1× PBS, and add 1× PBS to prevent drying.

6. Induction Evaluation: Observe the chondrogenic staining effect under a microscope, capture images, and evaluate the induction. Successful induction is indicated by the blue-green staining of acidic mucopolysaccharides in the cartilage tissue with Alcian blue.

NOTE: The chondrogenic differentiation level of mesenchymal stem cells varies depending on cell type, donor source, culture conditions, passage number, cell state, and differentiation time.