病院・クリニック向けPRP治療・幹細胞治療・再生医療支援（サポート）サービス／細胞塊スフェロイド三次元培養マイクロプレート「TASCL」

About This Manual

This manual is intended for use with TASCL 6WELL or TASCL DISH.
It describes the standard protocol, but please adjust as necessary depending on your application.
 
Note: TASCL MESH is designed to be used in combination with arbitrary equipment or gels (such as micro-well plates) and therefore does not have a dedicated manual. If necessary, please refer to this manual and adapt the procedures accordingly.
 
The following instructions are written with TASCL 6WELL in mind. When using TASCL DISH, replace “TASCL 6WELL” with “TASCL DISH” and replace “6-well plate” with “dish.”
 
Currently, TASCL series products are available for research use only. Administration of cell aggregates cultured with TASCL series products to humans is not permitted, except for treatments or clinical studies conducted in accordance with the Act on the Safety of Regenerative Medicine and related regulations.
If you are considering clinical studies in humans, please consult with us.
 
 

Materials to prepare inthe clean bench or the safety cabinet

- Packaged TASCL 6WELL / TASCL DISH
- Pipettes
- Pipette (100–1000 μl) or syringe for bubble removal by water flow (Not necessary if bubbles are removed using an incubator or centrifuge; see Step 2)
- Culture medium
- Sterile water
- Waste container for excess sterile water or medium
- If seeding cells: cell suspension (see Step 3)
- If harvesting aggregates: a container for the collected aggregates
 
 

Step 1. Check for Bubbles

This step is optional. You may skip directly to Step 2.
Add about 2 ml of sterile water onto the TASCL 6WELL, covering all microwells.
Check for the presence of bubbles inside the microwells.
If bubbles are present, they will reflect light, making the microwells appear whitish (see photo example).
If bubbles occupy more than ~5% of a microwell, proceed to Step 2.
If no bubbles are present or bubble removal is unnecessary, proceed to Step 3.
 

 
  

Step 2. Remove Bubbles from Microwells

Choose one of the following methods (A, B, or C):
 
A. Removal by Incubator
Keep TASCL with sterile water added (from Step 1) in a 37°C incubator for ~30 minutes.
After incubation, confirm bubble reduction.
If bubbles occupy less than 5% of the microwell, proceed to Step 3.
If bubbles remain, try method B or C.
 
B. Removal by Water Flow (Pipette/Syringe)
Add sterile water (up to half the insert height).
Use a pipette to aspirate sterile water and eject forcefully against the bubbles, removing them by water flow. (A disposable syringe may also be used.)
If bubbles are reduced to <5% of the microwell, proceed to Step 3. Otherwise, try method C.
 
C. Removal by Centrifugation
Place TASCL 6WELL/DISH (with sterile water added from Step 1) in a centrifuge with the lid on.
Centrifuge at 2000 RPM for ~5 minutes.
If bubbles are reduced to <5%, proceed to Step 3.
 
  

３．Prepare the cell suspension

For 621 microwells (TASCL 6WELL 600 / TASCL DISH 600):
Suspend desired number of cells per microwell × 621 in 0.5 ml medium.
 
For 1020 microwells (TASCL 6WELL 1000 / TASCL DISH 1000):
Suspend desired number of cells per microwell × 1020 in 0.5 ml medium.
 
Note: This method does not guarantee precise cell counts per microwell, but by distributing evenly across all microwells, similar numbers of cells will enter each microwell probabilistically.
 
 

Step 4. Cell Seeding

Aspirate and remove residual sterile water from microwells and outside the insert (6-well plate).
 
Rinse both microwells and plate with medium, then aspirate and remove.
Add ~2 ml of fresh medium into the microwells.
 
Dispense the prepared cell suspension evenly over the microwells, and allow cells to settle onto the mesh bottom.
 
Since sedimentation depends on cell type and medium density, it is recommended to confirm under an inverted microscope.
In most cases, cells will settle within 30 minutes.
 

Step 5. Addition of Medium

Add 3.0 ml of medium to the space outside the culture insert (between insert and 6-well plate).
 
To prevent drying of microwell surfaces by evaporation, ensure that the liquid levels inside the microwells and outside the insert are the same height. Adjust by gradually adding medium both inside and outside.
When medium volume decreases due to evaporation, replenish in the same manner.
 
Ensure the liquid level outside the insert is at least equal to that inside the microwells.
 
If the outside level is lower, a siphon effect may cause microwell medium to flow out during long-term culture.
Pre-warm medium to 37°C in an incubator (~10 minutes) if necessary.
 

Step 6. Medium Exchange

When exchanging medium, normally replace only the outside medium (between insert and 6-well plate).
 
Aspirate and replace all medium from the plate bottom via the gap between the insert and plate using a pipette.
 
Adjust volumes so that the liquid level inside the microwells matches the level outside.

Medium inside TASCL will gradually be exchanged with fresh medium through the porous insert membrane.
 

Step 7. Collection of Cell Aggregates

To harvest aggregates, add sufficient medium onto TASCL and gently pipette to detach aggregates, then collect together with medium.
 
Note: Aggregates may remain near the corners of square microwells. In such cases, repeat pipetting with added medium to collect aggregates.
 
Alternative methods if pipetting does not work:
 
- If aggregates adhere to TASCL: grasp TASCL with forceps, detach it from the insert, and release aggregates using pipette water flow into a 6-well plate or centrifuge tube containing buffer.
 
- If aggregates adhere to the culture insert surface: grasp TASCL with forceps, detach it from the insert, and scrape aggregates off the insert surface using a scraper.
 

 
 
End of manual.
For inquiries, please contact:
 
 
Revised on Sep. 13, 2025