Technical library

For most PromoCell Normal Human Cells, coated culture vessels are not required. Cells are usually cultured successfully on standard tissue‑culture‑treated plastic.
However, coating is required for certain cell types and applications. Since extracellular matrix (ECM) coatings can influence cellular metabolism and gene expression, the same coating should be used consistently within an experimental series.
You should always consult the respective Instruction Manual to confirm whether coating is required and which matrix is recommended.
 
When is coating required?

• Mesenchymal Stem Cells (MSC) (C‑12974/C‑12971/C‑12977): Require fibronectin, vitronectin, or collagen type I when cultured in MSC Growth Medium XF (C‑28019) and/or during differentiation with PromoCell MSC Neurogenic (C-28015), Adipogenic (C-28016), or Osteogenic (C-28013) Differentiation Media.
• Human monocyte‑derived macrophages (C‑12914/C‑12916/C‑12915/C‑12917): Must be seeded on fibronectin‑ or vitronectin‑coated vessels when using M1/M2 Macrophage Generation Media XF.
• Osteoblast mineralization assays: Collagen type I‑coated plates are recommended when using Osteoblast Mineralization Medium (C‑27020).

During quality control, PromoCell does not assess the presence or maintenance of specific stem cell markers in tumorspheres cultured in 3D Tumorsphere Medium XF (C‑28070).
 
Functional approach used instead
PromoCell applies a functional assay concept by culturing tumorspheres over serial passages in C‑28070.
Tumorsphere formation requires key biological properties such as:

• Anoikis resistance
• Self‑renewal capacity

These functional characteristics are considered hallmarks of cancer stem cells (CSCs) or cancer‑initiating cells (CICs).
Therefore, the ability to form and maintain tumorspheres over multiple passages functionally indicates the presence of CSCs/CICs, without relying on predefined marker expression.
 
Scientific advantage
3D tumorsphere culture enables researchers to:

• Study CSC biology in a functional context
• Avoid dependence on CSC marker panels, which can vary between tumor types and experimental systems

The abbreviations hMDM‑GMCSF(-) and hMDM‑MCSF(-) describe the origin, polarization, and activation state of the cryopreserved macrophages.
 
Meaning of the abbreviations
hMDM‑GMCSF(-):
hMDM: human monocyte‑derived macrophages
GM‑CSF: cells were differentiated (polarized) using GM‑CSF
(-): cells are not activated
→ These cells represent non‑activated M1‑polarized macrophages
hMDM‑MCSF(-):
hMDM: human monocyte‑derived macrophages
M‑CSF: cells were differentiated (polarized) using M‑CSF
(-): cells are not activated
→ These cells represent non‑activated M2‑polarized macrophages
 
Culture and activation

• Seed the macrophages in fibronectin‑coated tissue culture vessels
• After plating, they form biologically functional adherent cultures and can be maintained for several weeks
• Optional, user‑defined activation can be performed after seeding, depending on the experimental setup

Further details on polarization and activation options are provided in the Application Note "Standardized culture of assay-ready and fully functional human primary macrophages".

Yes. The following human cancer cell lines have been tested at PromoCell and shown to form tumorspheres when cultured in 3D Tumorsphere Medium XF (C‑28070):

• U‑87 MG
• MCF‑7
• MDA‑MB‑231
• HT‑29
• CT26
• HT1080
• HepG2
• A‑549
• Panc‑1
• LNCaP

In addition, customer feedback indicates successful tumorsphere formation for the following cell lines:

• A‑431 (human epidermal carcinoma)
• 4T1 (mouse mammary carcinoma)
• HCT‑116 (human colorectal carcinoma)
• Capan‑1 (human pancreatic adenocarcinoma)
• PC3 (human prostate cancer)
• C4-2B (human osteotropic prostate cancer)
• NCI‑H23 (human lung adenocarcinoma)
• IMR‑32 (human neuroblastoma)
• A818‑6 (human pancreatic ductal adenocarcinoma)
• HEK293 (human embryonic kidney)
• Calu‑1 (human non‑small‑cell lung cancer)

For experimental details, please refer to the Application Note "Tumorsphere culture of cancer stem cells". 

Yes. Based on customer feedback, PromoCell MSC Growth Medium 2 (C‑28009) has been successfully used to culture rat mesenchymal stem cells (MSCs).
According to this feedback, rat MSCs grow well in this medium and show good viability. Please note that this application is based on customer experience and has not been systematically validated by PromoCell.

Both Human Dermal Lymphatic Endothelial Cells (HDLEC) and Human Dermal Blood Endothelial Cells (HDBEC) express the pan‑endothelial marker CD31 (PECAM‑1).
 
HDLEC (C‑12216, C‑12217)
- CD31 (PECAM‑1): positive
- Dil-Ac-LDL uptake: positive
- Podoplanin: positive
Note: Podoplanin is a transmembrane glycoprotein involved in lymphatic vessel formation and is a characteristic marker of lymphatic endothelial cells.
 
HDBEC (C‑12211, C‑12225)
- CD31 (PECAM‑1): positive
- Dil-Ac-LDL uptake: positive
- Podoplanin: negative

Yes. Based on customer feedback, PromoCell 3D Tumorsphere Medium XF (C‑28070) has been successfully used for tumorsphere formation with mouse cell lines.
Please note that this application is based on customer experience and has not been systematically validated by PromoCell.

Phenol red is commonly used as a pH indicator in cell culture media. However, it has been shown to exhibit weak estrogen‑like (estrogenic) activity.
 
Use cases for phenol red–free media
Phenol red–free basal media are therefore recommended when:

• studying steroid hormone action, particularly estrogen signaling
• working with estrogen‑responsive cells

This estrogenic effect of phenol red was described by Berthois et al. (1986).
In addition, phenol red can interfere with analytical readouts, especially:

• colorimetric assays
• measurements where optical background or absorbance may affect data quality

The exact formulation of PromoCell basal media is proprietary information and is not publicly disclosed.
If you require information on the concentration of a specific component for your experiments, please contact PromoCell Scientific Support, who can advise you on a case‑by‑case basis.
 

The qualitative and quantitative composition of PromoCell supplements is provided:

• on the PromoCell website, and
• in the data sheets of the respective specialized media, where applicable.

If no detailed composition is specified, the formulation of the supplement is proprietary and confidential.