TC Protector/TC保护剂是一种细胞冷冻保护剂(Cell Cryoprotective agent)，不含任何动物源性成分或蛋白质，是化学定义的。可用于需要添加血清的细胞、无血清的细胞和无蛋白质的细胞。

TC Protector适用于细胞系细胞和正常细胞，以及ES细胞，成体干细胞和人iPS细胞。不粘稠，也不会形成气泡。因此，具有极好的可用性。

特征

• 根本不含动物源性成分，是化学定义的。

• 不含任何血清或蛋白质。

• 在冰箱中储存，取出后可立即使用。

• 粘性很小，可用性极好。

• 不需要特殊设备，例如程序冷冻机。可以直接存储于−80ºC。

冷冻方法

1. In case of adhesive cells, media should be changed at 80% confluence and be frozen on the next day.

In case of floating cells, media change (or media supplementation) is conducted and should be frozen on the next day.

*In order to freeze cells in a good condition, it is important to use cells from the logarithmic growth phase.

*Cells that have reached confluence or overgrown result in a decreased survival rate after freezing.

2. Cells are harvested according to the standard procedure and are centrifuged at 1,500 rpm for 1 minute.

3. It is diluted with TC Protector to achieve 5×105～1×107 cell/mL.

*Cell suspension is maintained in ice water when cell counts are measured.

*Since the appropriate cell number varies depending on cells, a pilot study must be conducted in advance. In general, 1×106 cells/mL is the target cell count.

4. Cells are stored at −80ºC in a Deep Freezer and are transferred into liquid nitrogen on the next day.

解冻方法

Ampules are placed in a warm water bath at 37ºC, and it is quickly thawed until it forms small ice mass.

Cell suspension is collected into a spitz tube and about 10× media is added. Then, it is centrifuged at 1,000–1,500 rpm for 1–2 minutes.

Cell pellet is checked and supernatant is discarded. Cells are seeded in accordance with the standard method.