Primary Human Preadipocytes & Growth Media

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

Preadipocyte/Adipocyte Cell Culture System

Adipose tissue is essential for energy storage and metabolic homeostasis of the body. Human White Preadipocytes (HWP) are self-renewing progenitors of mature differentiated adipocytes and can be found as a constitutive subpopulation in adipose tissue in adults.

PromoCell^® Human White Preadipocytes are produced from normal human subcutaneous and visceral adipose tissue from varied anatomical sites (lot-specific source information is available on request). Differentiation of HWP into mature adipocytes can be performed using the PromoCell^® Preadipocyte Differentiation Media system. Shortly after isolation, all Human White Preadipocytes are cryopreserved at passage 2 (P2) using proprietary serum-free freezing medium Cryo-SFM. Each cryovial contains at least 500,000 viable cells after thawing. Human White Preadipocytes are also available as proliferating cell cultures that are generated from cryopreserved cells that have been thawed and cultured for three days.

• Cultivation Protocol for Human White Preadipocytes

A range of culture media have been formulated for the undifferentiated expansion of human preadipocytes, differentiation of preadipocytes into mature adipocytes, and in vitro cultivation of human mature adipocytes. These media are optimized for growing primary human cells, but may also be used to cultivate murine preadipocytes/adipocytes.

• Preadipocyte Growth Medium - low-serum medium developed for the undifferentiated expansion of human preadipocytes.
• Preadipocyte Differentiation Medium - serum-free medium to induce the differentiation of preadipocytes into mature adipocytes.
• Adipocyte Nutrition Medium – media for in vitro cultivation of mature adipocytes.

All Preadipocyte/Adipocyte Media are available in two formats: Medium (ready-to-use) and Medium Kit. The Medium (ready-to-use) option consists of a 500 mL bottle of Basal Medium and one vial of SupplementMix.  The Medium Kit consists of a 500 mL bottle of Basal Medium and the SupplementPack (a set of individual vials with pre-measured supplements) to provide the user complete control over the media formulation. Adding the SupplementMix or the SupplementPack to the Basal Medium constitutes the complete medium.  The Basal Medium (with or without phenol red), SupplementMix, and SupplementPack are also available for individual purchase.

Preadipocyte/Adipocyte Media and Supplementation Details

The Preadipocyte Growth Medium contains all the growth factors and supplements necessary for the optimal growth of undifferentiated human preadipocytes. The Preadipocyte Differentiation Medium is a serum-free medium containing all supplements for effectively inducing the differentiation of preadipocytes into mature adipocytes, including ciglitazone. The Adipocyte Nutrition Medium is a low-serum medium containing all the growth factors and supplements necessary for the maintenance of mature adipocytes. These media do not contain antibiotics or antimycotics and are formulated for use in an incubator with an atmosphere of 5% CO₂.

Table 1. Final Preadipocyte/Adipocyte Media supplement concentrations (after addition to medium)

Component	Preadipocyte Growth Medium	Preadipocyte Differentiation Medium	Adipocyte Nutrition Medium
Fetal Calf Serum	0.05 mL / mL	—	0.03 mL / mL
Endothelial Cell Growth Supplement	0.004 mL / mL	—	—
Epidermal Growth Factor (recombinant human)	10 ng / mL	—	—
Hydrocortisone	1 μg / mL	—	—
d-Biotin	—	8 μg / mL	8 μg / mL
Insulin (recombinant human)	—	0.5 μg / mL	0.5 μg / mL
Dexamethasone	—	400 ng / mL	400 ng / mL
IBMX	—	44 μg / mL	—
L-Thyroxine	—	9 ng / mL	—
Ciglitazone	—	3 μg / mL	—
Heparin	90 μg / mL	—	—

Preparing Supplemented Media for Preadipocyte/Adipocyte Culture

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

Note: Light flocculation may be seen upon thawing the supplements containing ECGS. This does not affect reagent activity. Optionally, the precipitate can be removed by centrifugation under sterile conditions.

Instructions for Use of Preadipocyte Differentiation Medium and Adipocyte Nutrition Medium

• For differentiating human preadipocytes, culture the cells in Preadipocyte Growth Medium to confluency.
• Replace the Growth Medium with Preadipocyte Differentiation Medium and culture for 72 hours.
• After 72 hours, replace the Preadipocyte Differentiation Medium with Adipocyte Nutrition Medium to complete the differentiation process.
• Feed the cells every 2 – 3 days with fresh Adipocyte Nutrition Medium. Differentiation to mature adipocytes is typically achieved after 12 – 14 days.

Storage and Stability of Preadipocyte/Adipocyte Products

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

Preadipocyte/Adipocyte Culture System Quality Control

Human White Preadipocytes (HWP) Quality Control

Rigid quality control tests are performed for each lot of Human White Preadipocytes.

• Tested for cell morphology, adherence rate, and cell viability.
• Tested for capacity to differentiate into mature adipocytes.
• Growth performance is tested through multiple passages up to 10 population doublings (PD) under culture conditions without antibiotics and antimycotics.
• Tested for the absence of HIV-1, HIV-2, HBV, HCV, HTLV-1, HTLV-2 and microbial contaminants (fungi, bacteria, and mycoplasma).
• A detailed certificate of analysis (CoA) for each lot is available for download.

Table 2. Human White Preadipocytes Specifications

Product	Plating Density	Passage after Thawing	Marker	Population Doublings
Human White Preadipocytes (HWP) subcutaneous	5,000 cells per cm²	P2	Differentiation tested	> 10
Human White Preadipocytes (HWP) visceral	5,000 cells per cm²	P2	Differentiation tested	> 10

Preadipocyte/Adipocyte Media Quality Control

All lots of Preadipocyte/Adipocyte Media are subjected to comprehensive quality control tests using primary human white preadipocytes. Each lot is checked for growth-promoting activity, adherence promotion, and typical morphology of the tested preadipocytes. In addition, the Preadipocyte Differentiation Medium/Adipocyte Nutrition Medium are tested for the capacity to induce the terminal differentiation of preadipocytes into mature adipocytes. Approved in-house lots of Preadipocyte/Adipocyte Media are used as reference.  All lots of media are rigorously tested for the absence of microbial contaminants (fungi, bacteria, mycoplasma).

Intended Use for Primary Preadipocyte/Adipocyte Culture Systems

These products are for in vitro use only and not for diagnostic or therapeutic procedures. For safety precautions please see appropriate MSDS.

Note:  Due to their low serum content or the absence of serum, PromoCell media are not suitable for 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.