Technical library

Freshly isolated monocytes readily attach when cultured in Monocyte Attachment Medium (C‑28051), whereas cryopreserved CD14⁺ monocytes (C‑12909) largely lose this attachment capability after thawing.
Monocyte attachment is important not only for establishing an adherent culture but also as a key step in the differentiation process from monocytes to macrophages.
As a result, cryopreserved CD14⁺ monocytes generally show reduced macrophage differentiation efficiency, leading to lower yields and less complete differentiation compared to freshly isolated monocytes.
For optimal macrophage generation, we therefore recommend using freshly isolated PBMC or monocytes in combination with Monocyte Attachment Medium (C‑28051).

Yes. Both the distinct odor and the color change are normal characteristics of Cryo-SFM Plus and are caused by the antioxidant technology used in the freezing medium. These properties do not affect cell performance and pose no risk to the user or the cells.
 
Background: Cryo-SFM Plus contains a proprietary antioxidant system designed to enhance post-thaw cell viability, attachment, and growth.
 
Please note:

• Frozen Cryo-SFM Plus appears yellow
• After thawing, the medium turns orange
• Cryo-SFM Plus does not turn pink

No. None of the media in PromoCell's Cancer Media Toolbox contain ROCK inhibitors or other small-molecule inhibitors. This includes:

• Primary Cancer Culture System (PCCS, C‑28081)
• 3D Tumorsphere Medium XF (C‑28070)
• Cancer Cell Line Medium XF (CCLM XF, C‑28077)

No. Although both Keratinocyte Growth Medium 2 and Keratinocyte Growth Medium 3 are serum-free, they differ in more than just the presence or absence of Bovine Pituitary Extract (BPE).
Keratinocyte Growth Medium 2 contains BPE and is designed for the cultivation of epidermal keratinocytes. Keratinocyte Growth Medium 3 is specifically formulated to be BPE-free and features a different supplement composition.
As a result, simply omitting BPE from Keratinocyte Growth Medium 2 does not generate a medium equivalent to Keratinocyte Growth Medium 3.
 
Please note: Keratinocyte Growth Medium 3 offers a higher level of standardization by eliminating BPE and supports the expansion of fast‑proliferating, homogeneous keratinocyte cultures. However, according to PromoCell's media classification, it is not considered a "defined" medium.

Yes, SkMC (C‑12530) can be cultured in Skeletal Muscle Cell Growth Medium (C‑23060) without using differentiation medium. Under these conditions, the cells remain as proliferating myoblasts.
These proliferating cells can be used for studies focusing on the induction or inhibition of cell proliferation.
However, to study the function of differentiated skeletal muscle cells (multinucleated myotubes), it is necessary to switch to Differentiation Medium (C‑23061) once the cells reach 60–80% confluency.
A detailed differentiation protocol is provided in the Instruction Manual for SkMC media.

HUVEC (both single donor and pooled, including C‑12203) can be cultured in 

• Endothelial Cell Growth Medium (C‑22010) as well as in
• Endothelial Cell Growth Medium 2 (C‑22011).

Both media support robust growth of HUVEC; however, they differ slightly in their supplement composition.
 
At PromoCell, Endothelial Cell Growth Medium (C‑22010) is the standard medium used for expansion of HUVEC cultures and for Quality Control testing.
In addition, PromoCell also offers HUVEC-pooled isolated in Endothelial Cell Growth Medium 2 (C‑12208). These cells are the preferred choice when Endothelial Cell Growth Medium 2 is used as the routine culture medium.
 

The viability of detached macrophages depends largely on whether the cells can be harvested using Macrophage Detachment Solution alone or whether additional mechanical detachment is required.

• If the macrophages can be detached using Macrophage Detachment Solution alone (i.e. without scraping), the expected viability is typically 50-80%.
• If a cell scraper must be used in addition to the Macrophage Detachment Solution, viability is usually below 50% and may be as low as ~30%.

Please note: Macrophages are highly adherent cells, and some loss of viability during detachment is expected. To maximize recovery and viability, mechanical stress should be minimized whenever possible.

Based on feedback received from users, coating the culture vessel with Fibronectin or Poly‑L‑Lysine is recommended for calcification studies with vascular smooth muscle cells.
Users have reported successful calcification of PromoCell Human Aortic Smooth Muscle Cells (HAoSMC) cultured on Fibronectin‑ or Poly‑L‑Lysine‑coated culture vessels and treated with Osteoblast Mineralization Medium (C-27020) in a 21‑day calcification assay.
In addition, successful calcification has been reported for Human Coronary Artery Smooth Muscle Cells (HCASMC) cultured on Fibronectin‑coated culture vessels.
 
Please note: These observations are based on user feedback and have not been independently validated by PromoCell. 

Cryopreserved human macrophages require a specialized thawing protocol because they are particularly sensitive to freeze-thaw stress.
During the recovery phase, the cells are incubated in cold medium (2-8°C) for 20 minutes before seeding. This gradual warming process allows the cells time to recover and restore membrane integrity, resulting in improved cell viability and attachment.
Based on our experience, this recovery step significantly enhances the adhesion efficiency of cryopreserved human macrophages after thawing.

Please note: For detailed thawing instructions, please refer to the corresponding Instruction Manual.

Yes. With a few exceptions, established cancer cell lines can be cultured in the Primary Cancer Culture System (PCCS, C‑28081) to enrich cancer stem cells (CSCs) for downstream research applications.
Please note: During the initial passages, cell growth may be slower than in conventional culture media.
 
Background
Established cancer cell lines are capable of indefinite proliferation under standard culture conditions. However, they typically contain only a small fraction of CSCs or CSC‑like cells (approximately 1–20%), while the majority of cells are more differentiated.
When cultured in PCCS, the proportion of CSC‑like cells increases, whereas the proportion of differentiated cells decreases. Since CSCs generally proliferate more slowly than the bulk tumor cell population, a reduction in overall growth rate may be observed during the adaptation phase.
In addition, cells may undergo morphological changes and appear smaller and more compact, reflecting enrichment of a more stem‑like and tumorigenic subpopulation.
 
Alternative medium
Unlike PCCS, Cancer Cell Line Medium XF (CCLM XF, C‑28077) is designed to support the growth of established cancer cell lines without enriching for CSCs. While CSCs can be maintained in CCLM XF, the medium does not selectively favor them. As a result, cell growth rates are typically comparable to those observed in conventional culture media.
 
Our Scientific Support team will be happy to discuss whether PCCS or CCLM XF is the more suitable option for your specific research application.