Primary Human Smooth Muscle Cells and Growth Media

• Human Smooth Muscle Cell Culture System
• Smooth Muscle Cell Culture System Quality Control
• Related Products

Human Smooth Muscle Cell Culture System

Smooth muscle tissue is found in the tunica media layer of large and small blood vessels and in the walls of hollow organs like the bladder and the uterus. All Smooth Muscle Cells (SMC) possess the same types of filaments, but depending on the tissue of origin, differ significantly in mechanical and physiological properties.

PromoCell^® Human Smooth Muscle Cells (SMC) are isolated from normal human tissue of diverse origins including vascular smooth muscle cells from sources like the aorta, the coronary artery, the pulmonary artery, the umbilical artery.  SMC isolated from the trachea, the bronchi, and the uterus are also available. Shortly after isolation, all human Smooth Muscle Cells are cryopreserved at passage 2 (P2) using proprietary, serum-free freezing medium, Cryo-SFM (C-29910). Each cryovial contains more than 500,000 viable cells after thawing. Human Smooth Muscle Cells are also available as proliferating cell cultures generated from cryopreserved cells that have been thawed and cultured for three days.

The Smooth Muscle Cell Growth Medium 2 was developed for the in vitro cultivation of smooth muscle cells from blood vessels and hollow organs. The medium is optimized for growing primary human cells, but can also be used to culture bovine, porcine, dog, murine, and rat smooth muscle cells. The Smooth Muscle Cell Media is available in two options: “ready-to-use” and “kit”. The “ready-to-use” option consists of a 500 mL bottle of Basal Medium and one vial of SupplementMix. The Smooth Muscle Cell Medium Kit consists of a 500 mL bottle of Basal Medium and the SupplementPack (a set of individual vials with pre-measured supplements), allowing the user full control over the media formulation. Add the SupplementMix or the SupplementPack to Basal Medium to prepare the complete Smooth Muscle Cell Growth Medium. The Basal Medium (with or without phenol red), SupplementMix, and SupplementPack are also available for separate purchase.

Smooth Muscle Cell Growth Medium 2 is recommended for:

• Human Umbilical Artery Smooth Muscle Cells (HUASMC)
• Human Coronary Artery Smooth Muscle Cells (HCASMC)
• Human Pulmonary Artery Smooth Muscle Cells (HPASMC)
• Human Aortic Smooth Muscle Cells (HAoSMC)
• Human Bronchial Smooth Muscle Cells (HBSMC)
• Human Tracheal Smooth Muscle Cells (HTSMC)
• Human Uterine Smooth Muscle Cells (HUtSMC)

Smooth Muscle Cell Growth Medium 2 Supplementation Details

The Smooth Muscle Cell Growth Medium 2 contains all the growth factors and supplements necessary for the optimal growth of human smooth muscle cells, including epidermal growth factor, basic fibroblast growth factor, and insulin (see Table 1 for concentrations). Smooth Muscle Cell Growth Medium 2 does not contain antibiotics or antimycotics and is formulated for use in an incubator with an atmosphere of 5% CO₂.

Table 1. Final Smooth Muscle Cell Growth Medium 2 supplement concentrations (after addition to the medium).

Component	Final Concentration
Fetal Calf Serum	0.05 mL / mL
Epidermal Growth Factor (recombinant human)	0.5 ng / mL
Basic Fibroblast Growth Factor (recombinant human)	2 ng / mL
Insulin (recombinant human)	5 μg / mL

Preparing Supplemented Media for Smooth Muscle Cell Culture

• Thaw the Smooth Muscle Cell Growth Medium 2 SupplementMix or SupplementPack at 15 – 25 °C.
• Aseptically mix the supplement solutions by carefully pipetting up and down.
• Transfer the entire contents of each supplement to the Smooth Muscle Cell Basal Medium.
• Close the bottle and swirl gently until a homogeneous mixture is achieved.

Storage and Stability of Smooth Muscle Cell Products

• Upon arrival, immediately store the Smooth Muscle Cell Basal Medium at 4 – 8 °C in the dark; do not freeze the medium. Store the Smooth Muscle Cell Growth Medium SupplementMix or SupplementPack at -20 °C.
• If stored as directed, the products are stable until the stated expiration date.
• Once supplements are added to the Smooth Muscle Cell Basal Medium, the shelf life of the complete medium is six weeks at 4 – 8 °C. Do not freeze the complete medium.
• For use, prewarm only an aliquot of the complete Smooth Muscle Cell Growth Medium needed for single use, and keep the remaining medium refrigerated at 4 – 8°C.
• After delivery, cryopreserved cells should be stored in liquid nitrogen or seeded directly.
• Proliferating cells must be processed immediately.

Smooth Muscle Cell Culture System Quality Control

Human Smooth Muscle Cells Quality Control

Rigid quality control tests are performed for each lot of Human Smooth Muscle Cells.

• Analysis of cell morphology, adherence, and cell viability
• Flow cytometric analyses for cell-type specific markers such as smooth muscle alpha-actin
• Growth performance, as tested through multiple passages up to 15 population doublings (PD) under culture conditions without antibiotics and antimycotics
• Testing for the absence of viral (HIV-1, HIV-2, HBV, HCV, HTLV-1, HTLV-2) and microbial (fungi, bacteria, and mycoplasma) contaminants. For safety precautions, please refer to the appropriate safety data sheet (SDS).

A detailed certificate of analysis (CoA) for each lot is available for download.

Table 2. Smooth muscle cell specifications.

Product	Plating Density	Passage after thawing	Marker	Population Doublings
Human Aortic Smooth Muscle Cells (HAoSMC)	7,500 - 10,000 cells per cm²	P2	Smooth muscle α-actin+	> 15
Human Coronary Artery Smooth Muscle Cells (HCASMC)	7,500 - 10,000 cells per cm²	P2	Smooth muscle α-actin+	> 15
Human Pulmonary Artery Smooth Muscle Cells (HPASMC)	7,500 - 10,000 cells per cm²	P2	Smooth muscle α-actin+	> 15
Human Umbilical Artery Smooth Muscle Cells (HUASMC)	7,500 - 10,000 cells per cm²	P2	Smooth muscle α-actin+	> 15
Human Tracheal Smooth Muscle Cells (HTSMC)	7,500 - 10,000 cells per cm²	P2	Smooth muscle α-actin+	> 15
Human Bronchial Smooth Muscle Cells (HBSMC)	7,500 - 10,000 cells per cm²	P2	Smooth muscle α-actin+	> 15
Human Uterine Smooth Muscle Cells (HUtSMC)	7,500 - 10,000 cells per cm²	P2	Smooth muscle α-actin+	> 15

Smooth Muscle Cell Growth Medium Quality Control

All lots of Smooth Muscle Cell Growth Medium are subjected to comprehensive quality control tests using primary human smooth muscle cells. Each lot is checked for growth promoting activity, adherence rate, and typical morphology of the tested smooth muscle cells. Approved in-house lots of Smooth Muscle Cell Growth Media are used as a reference.

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

Intended use for Smooth Muscle Cell Culture system products.

PromoCell^® Primary Human Smooth Muscle Cells and Media are for in vitro use only and not for diagnostic or therapeutic procedures.

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