Product Highlight

• The culture medium has been filed with FDA DMF with filing No. of 37874
• The product is a serum-free culture kit for pure factor culture of CIK cells, suitable for CIK cells culture of umbilical cord blood samples.
• Standard operation for different samples: Provided that the initial PBMC inoculation conditions are controlled, the subsequent cultures can be carried out according to the recommended fluid replacement process.
• Stable culture results: This product has been on the market for 7 years, with highly reliable stability.
• It is used for the in vitro induction and amplification of CIK cells from umbilical cord blood samples and is NOT applicable to peripheral blood samples.
• It is applicable to sample volumes of 50 mL of umbilical cord blood and uses 2-3 Liters of culture medium.

Product Background

YOCON serum-free culture kit for pure factor culture of CIK cells is suitable for CIK cell culture of peripheral blood and umbilical cord blood samples. After 14-16 days of culture, the cell count reaches at least 5.0 billion, and the positive rate is over 20%. This product is composed of a basic medium and an induced expansion kit, to provide an extremely easy-to-use solution for CIK cell culture.

Cytokine-induced killer (CIK) cell therapy is a type of adoptive immunotherapy that due to its high proliferation rate and anti-tumor characteristics, is being investigated to treat various solid tumors. Since advanced colorectal cancer (CRC) has high mortality and poor survival rates, and the efficacy of chemotherapy and radiotherapy is limited in treatment, the application of CIK cell therapy in CRC has been evaluated in numerous studies. This review aims to summarize the clinical studies that investigated the safety and clinical efficacy of CIK cell therapy in CRC. Therefore, 1,969 enrolled CRC patients in the clinical trials, of which 842 patients received CIK cells in combination with chemotherapy with or without dendritic cell (DC) infusions, were included in the present review. Furthermore, the signaling pathways involved in CIK cell therapy and novel methods for improving migration abilities are discussed.

The total number of 4-6 billion cells can be obtained after 14 days of culture in a 2 L system, and the proportion of CD3+CD56+ is stable at 10% to 20%.
Pure factor induction, with no trophoblast cells, with the simple and easy culturing method, when combined with the newly upgraded basic culture medium of immune cells, significantly improved the continuous expansion ability of cells after 7 consecutive days of culturing, and the continuous support ability of cells expanded from 2 L system to 3 L or even 4 L system after 14 days.

Fluid Replacement Procedure 1: Final volume of 2 liters.

Day	Culture Consumables	Culture Medium	Fluid replacement volume (mL)	Total Vol (mL)	Plasma proportion (%)	Plasma volume (mL)
0	1x T75 flask	Activation medium	30	30	10	3
3	1x T75 flask	Activation medium	30	60	5	1.5
5	1x T75 flask	Activation medium	140	200	5	7
7	1x culture bag	Expansion medium	200	400	1	2
9	2x culture bag	Expansion medium	400	800	0	0
11	2x culture bag	Expansion medium	800	1600	0	0
13	2x culture bag	Expansion medium	400	2000	0
16-18	2x culture bag	Harvest cells

Fluid Replacement Procedure 2: Final volume of 3 liters.

Day	Culture Consumables	Culture Medium	Fluid replacement volume (mL)	Total Vol (mL)	Plasma proportion (%)	Plasma volume (mL)
0	1x T75 flask	Activation medium	30	30	10	3
3	1x T75 flask	Activation medium	30	60	5	1.5
5	1x T75 flask	Activation medium	140	200	5	7
7	1x culture bag	Expansion medium	200	400	1	2
9	2x culture bag	Expansion medium	400	800	0	0
11	2x culture bag	Expansion medium	800	1600	0	0
13	2x culture bag	Expansion medium	1400	3000	0
16-18	2x culture bag	Harvest cells

Special Instructions

1. Isolation of PBMC
   Two points should be particularly noted when isolating mononuclear cells. First, before isolating mononuclear cells, the blood and all reagents should be pre-warmed to 20°C and centrifuged at room temperature. Second, PBMC should be isolated within 8 hours after blood collection.
2. Inoculation density
   The recommended inoculation density for fresh umbilical cord blood mononuclear cells is 2×10⁶/mL, while that for cryopreserved umbilical cord blood mononuclear cells is 3×10⁶/mL. An inoculation density that is too low or too high will have an impact on the final number of harvested cells and the purity of CIK cells.
3. Reagent preservation
   The factors in the CIK cell kit are prohibited from being repeatedly frozen and thawed, otherwise their activity will be reduced, resulting in a lower positive rate.