Technical library

Yes, it is possible. Short protocol: Plate human osteoblasts in Osteoblast Mineralization Medium on collagen I coated TC vessels. Incubate the cells for 17-21 days and change the medium every third day. Be careful not to disturb the cell monolayer. Fix the cells. The calcium deposition can be visualized by von Kossa or Alizarin Red staining. More detailed information on osteoblast mineralization and Alizarin Red S staining can be found in the attached Application Note.

Our HCASMC (C-12221) are isolated from the large arteries, i.e. from

• Right coronary artery
• Left main coronary artery
• Circumflex coronary artery and
• Left anterior descending coronary artery.

Both, trypsin and accutase represent mixtures of different proteolytic enzymes. Trypsin is prepared from porcine pancreas, accutase from invertebrates. Accutase can replace trypsin for the detachment and dissociation of anchorage-dependent cells from surfaces and can also be used on suspension cells to reduce clumping in preparation for counting. The advantages of accutase over the traditional trypsin treatment are that it is more gentle and less damaging to cells (leading to increased viability) and does not contain any mammalian or bacterially derived proteins.

Accutase is more thermolabile than trypsin and usually doesn't require an inactivation step.

We recommend to use the trypsin as well as the other detach solutions at room temperature to avoid overtrypsinization and irreversible cellular damage.

Upon arrival, the basal media should be stored between 4°C and 8°C, the supplements at -20°C.

Please do not freeze our cell culture media. Freezing can lead to irreversible precipitation of media components and the quality can no longer be assured.

Yes, you can use accutase to detach Normal Human Cells. Accutase acts very gently on the cells. Cell membranes and surface epitopes will not be harmed. It is therefore mostly used for applications that require unchanged surface markers, e.g. for flow cytometry, or for detachment of very sensitive cells.

The PromoCell Trypsin/EDTA (ready-to-use) and the PromoCell TNS solution are both based on HepesBSS. Therefore, it is best to use HepesBSS to wash the cells prior to trypsinization.
However, PBS can also be used.

We recommend using the DetachKit (C-41200) for subculturing our normal human cells. 

It was designed for the safe and efficient detachment of primary human cells in routine subculturing and consists of 3 components: HEPES BSS (HEPES buffered Balanced Salt Solution), Trypsin/EDTA Solution and TNS (Trypsin Neutralizing Solution). All 3 components are ready-to-use.

Please note: Many of our culture media have low serum content or no serum at all. These media are not suitable for inactivating trypsin during subculture.

The population doubling time or generation time (t_g) is usually calculated during the logarithmic phase of growth. It specifies the time (t) in hours needed by the culture to double its cell number.

t_g = t / n

t = time (hrs) between 2 cell counts

n= number of population doublings