Primary Human Chondrocytes and Chondrocyte Growth Media

These products are available in the US, Canada, and select European countries.

• Chondrocyte Culture System
• Cultivation Protocol for Human Chondrocytes
• Quality Control for Human Chondrocytes and Chondrocyte Growth Media
• Chondrocyte Culture Materials

Human Primary Chondrocyte Culture System

In the human body, articular cartilage covers the bones in the major joints, such as knee and hip joints. It is composed solely of chondrocytes, which are specialized cells that produce and maintain the extracellular matrix of cartilage including collagen (mostly type II) and proteoglycans. In vitro-differentiated chondrocytes express collagen type II; however, this expression declines as chondrocytes dedifferentiate during prolonged cultivation.

PromoCell^® Human Chondrocytes are isolated from knee and hip joint cartilage, and cryopreserved shortly thereafter, at passage 2 (P2). Primary chondrocytes are isolated and cultured using proprietary, serum-free freezing medium, Cryo-SFM. Each thawed cryovial contains at least 500,000 viable cells. Human Chondrocytes are also available as proliferating cell cultures generated from cryopreserved cells that have been thawed and cultured for three days.

• Cultivation Protocol for Human Chondrocytes

Chondrocyte Growth Medium was developed for the in vitro cultivation of primary human chondrocytes. This medium is optimized for growing primary human cells, but can also be used for growing primary porcine and rat chondrocytes. The Chondrocyte Growth Medium includes a 500 mL bottle of Basal Medium and one vial of SupplementMix. Adding the SupplementMix to the Basal Medium constitutes the complete Chondrocyte Growth Medium. Basal Medium (with or without phenol red) and the SupplementMix are also available for separate purchase.

Primary Chondrocyte Growth Media Supplementation Details

The Chondrocyte Growth Medium includes all the components necessary for the optimal growth of human chondrocytes. Chondrocyte Growth Medium does not contain antibiotics or antimycotics and is formulated for use in an incubator with 5% atmospheric CO₂.

Chondrocyte Growth Medium supplement concentrations
(after addition to the medium)

Component	Final Concentration
Fetal Calf Serum	0.1 mL / mL

Preparing Supplemented Medium for Primary Chondrocytes

• Thaw the Chondrocyte Growth Medium SupplementMix at 15 – 25° C.
• Aseptically mix the supplement solutions by gently pipetting up and down.
• Transfer the entire contents of each desired supplement to the Chondrocyte Basal Medium.
• Close the bottle and swirl gently to achieve a homogeneous mixture.

Storage and Stability of Human Chondrocytes and Chondrocyte
Growth Medium

• Upon arrival, immediately store the Chondrocyte Basal Medium in the dark at 4 – 8 °C; do not freeze. Store the Chondrocyte Growth Media SupplementMix at -20 °C.
• If stored properly, the products are stable until the expiration date stated on the label.
• Once the supplement is added to the Chondrocyte Basal Medium, the shelf life of the complete medium is six weeks at 4–8 ° C. Do not freeze the complete medium.
• To use, prewarm just the required volume of the complete Chondrocyte Growth Medium and keep the remaining medium refrigerated at 4 – 8° C.
• Upon arrival, seed cells directly, or preserve in liquid nitrogen.
• Proliferating cells must be processed immediately.

Quality Control for Human Chondrocytes and Chondrocyte Growth Media

Human Chondrocytes Quality Control

The following rigid quality control tests are performed on each lot of Human Chondrocytes:

• Cell morphology, adherence rate, and cell viability testing
• Testing for the ability to differentiate in 3D spheroids
• Growth performance testing through multiple passages up to 10 population doublings (PD) under culture conditions without antibiotics and antimycotics
• Testing for the absence of HIV-1, HIV-2, HBV and HCV, and microbial contaminants (fungi, bacteria, and mycoplasma)

A detailed certificate of analysis (CoA) for each lot can be downloaded.

Human Chondrocytes Specifications

Product	Plating Density	Passage after Thawing	Marker	Population Doublings
Human Chondrocytes	10,000 - 20,000 cells per cm²	P2	Differentiation tested	> 10

Primary Chondrocyte Growth Medium

All lots of Chondrocyte Growth Medium are subjected to comprehensive quality control testing. These tests check for growth promoting activity, adherence rate, and typical morphology of primary human chondrocytes. Approved in-house lots of media are used as a reference. Additionally, microbial testing ensures that all lots of Chondrocyte Growth Media are free of contaminants (fungi, bacteria, mycoplasma).

Intended Use for Human Chondrocytes and Chondrocyte Growth Media

Human Chondrocytes and Chondrocyte Growth Media are intended for in vitro use only and not for diagnostic or therapeutic procedures. For safety information please see appropriate MSDS.

Note: PromoCell^® media have low or zero serum content and are not suitable for trypsin neutralization (when subculturing the cells, for example). Instead we recommend using the DetachKit (C-41200, C-41210, C-41220), which contains HEPES BSS, Trypsin/EDTA and Trypsin Neutralizing Solution.

Human Chondrocyte Culture Materials