Tag Archives: consumables

HBVPs ready and on the go!

I am now well into culturing thanks to Jo-Maree’s expert guidance. I am happy to say that it is all coming back to me and I actually feel pretty confident navigating the labyrinthian lab layout and doing routine cell culture.

I also have my own stock of HBVPs in the Stem Cell lab. As you can see they are quite spindly and form little ‘star clusters’ as they grow:


Brightfield Microscope image of HBVP cells (P5 – passage no. 5). These cells were passaged on 9/9/21.

These cells grow in Complete Pericyte Medium (CPM) – Pericyte Medium supplemented with FBS, Antibiotics and Growth Supplements. When keeping a lab journal, it is important to record the Lot no. for all items. This allows researchers to track any variations between batches. It is also vital to record the opening date.

Complete Medium

Complete Pericyte Medium – prepared on 9/9/21

The HBVPs grow in standard T75 Tissue Culture flasks in 15mL of CPM (with cell type, passage number, date of passage and researcher initials listed on the vessel).

HBVP cells

HBVP cells (P4 – passage 4) in T75 Flask. 

As part of the experimentation process, I prepared  three 90mm glass Petri dishes.

Protection from AccidentsI engraved the base of two dishes with a ripple pattern to see if the engraving would impact on the growth/adherence of the cells. Inspired by the awesome work of Whitefeather Hunter (and my previous collaborative work with sigils), one of these dishes also had an engraved symbol to aid protection from accidents.

As you can see below, the engraving looks quite rugged and sharp as it was done by hand. I am curious to see if the cells will grow on this area.

Engraved Glass

Brightfield microscope image of the engraved glass.

To sterilise the dishes for cell culture, they are placed into paper autoclave bags and sealed with autoclave tape.  They are then placed into an autoclave that steam sterilises the vessels at high temperature. The black lines on the tape indicate that the process was successful and the correct temperature was reached.

Autoclaved Petri Dish

Single Petri dish in autoclave bag – dated 26/8/21

Most cells do not adhere well to untreated plastic or glass surfaces. As such we added a Poly-L-lysine coating to aid cell adherence.


Poly-L-lysine solution.

The Poly-L-lysine solution was diluted with sterile H2O to make up 30mL total (10mL for each Petri Dish).


10mL of the Poly-L-lysine solution was added to each dish and then incubated for an hour.

Petri Dishes

After an hour, I removed the Poly-L-lysine solution and washed each dish twice with sterile H2O.  I passaged (split) the HBVPs and added approx 1 million cells (in 1mL media) to each Petri Dish and one T75 Flask (plus 14mL media).


Material supplies: Glass Petri Dishes

My primary lab contact is currently on leave so I am using the time to identify materials for experimentation. I am keen to grow and stain my cells on diverse materials (glass, porous and non-porous scaffolds etc). An easy start is to use glass Petri dishes with different coatings to encourage cell adherence. The use of coatings may also enable me to encourage cells to grow in particular patterns.

I’ve done a bit of searching via Researchgate and it seems that common surface coatings to encourage cell attachment to glass include:

  • Hyaluronic acid
  • Poly-D-Lysine – my current choice
  • Fetal Calf Serum
  • Bovine serum albumin
  • Gelatin
  • Fibronectin
  • Laminin
  • Collagen

I will ask for advice at the next lab meeting.

There were also suggestions to etch the glass surface with concentrated nitric acid and then wash and autoclave. At the UTAS School of Creative Arts and Media, we are fortunate to have access to a glass cutter and laser engraver. So instead of using acid, I will use the laser to score a design into the base of a large 150mm Petri dish.

Glass Petri Dish

Glass Petri dish by Lilly M via Wikimedia

Luckily Petri dishes are easy to purchase online. I just need to make sure the dishes are suitable for autoclave sterilisation (e.g.  borosilicate glass rather than soda lime glass).