NK Cell Large-Scale Culture Kit|Umbilical Blood|Yocon Cell

Umbilical Cord Blood NK Cell Large-Scale Culture Kit:A modular system utilizing defined cytokine cocktails (NK-A/B/C) for feeder-free NK cell expansion, delivering >90% viability and 70–90% purity at scales up to 5×10⁶ cells/mL.（ZL201510624087.4）
Immune Repair Culture Medium:A GMP-grade, animal-component-free medium that repairs damaged PBMCs via IL-2/IL-15 priming, reducing culture failure rates by 40–60% in suboptimal samples.
NK Cell Serum-Free Medium (Recombinant Human Albumin):A chemically defined, xeno-free medium sustaining >90% viability for 5+ days at 5×10⁶ cells/mL, optimized for closed-system bioreactor integration.
FEP Culture Bags (2L):Inert FEP polymer bags (transparent, -196°C to +137°C durable) enabling 20–40% higher yields than EVA via enhanced gas permeability and sterile, needle-free ports.
（Catalog Number：DMF）：（NC1011：37754；NC0102.H：37874；NC0102.F：40625；FP0002：041754）

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High Yield Efficiency

The primary advantage of our kit is its unparalleled ability to consistently achieve high yields of functional NK cells from umbilical blood, which is crucial for successful clinical and research applications.

Tailored Adaptability

Its unique modularity offers unmatched flexibility, enabling you to precisely tailor the culture process to specific blood volumes. This approach significantly minimizes waste and maximizes efficiency, optimizing your resource utilization.

Robust Cell Functionality

Through a carefully formulated culture environment, the kit ensures robust cell expansion while maintaining the vital functionality of the NK cells, pushing the boundaries of what's achievable in immune cell research and therapy

NK Cell Expansion Case

Sample	Transportation Duration	Cryopreserved	Anticoagulant Proportion	Inoculation Number	Final Volume	Culture Duration	Expansion Fold	Harvested Cells	NK Phenotype
1	Approximately 24h	Cryopreserved	21.5%	60 million	4L	26 days	253	15.2 billion	99.05%
2	Approximately 24h	Fresh	18.7%	60 million	6.3L	27 days	428	25.7 billion	98.54%
3	Approximately 24h	Cryopreserved	25.0%	20 million	6.3L	28 days	370	7.4 billion	98.67%
4	Approximately 24h	Fresh	20.0%	120 million	12.9L	29 days	408	48.9 billion	99.22%
5	Approximately 24h	Fresh	19.0%	60 million	6.12L	28 days	351	21 billion	97.52%

Product information

NK Large-Scale Culture Kit —— Standard Components

Product Name	Catalog Number	Application	Specification	Storage
Immune Cell Repair Culture Medium	NC0102.F	Used for early activation of NK cells	1 bottle, 200mL/bottle	2 ~ 8°C
Serum - free Culture Medium for NK Cells (Recombinant Human Albumin)	NC0102.H	In vitro culture of NK cells	6 bottles, 1L/bottle	2 ~ 8°C
NK Cell Induction and Expansion Kit (High - performance Version)	AN0104 - 2	YC00A: Used for coating culture flasks during initial culture	1 vial, 500μL/vial	-20°C
		YC00B: Added during initial culture	1 vial, 500μL/vial
		YC00C: Added to activation culture medium	1 vial, 200μL/vial
		YC005: Added to expansion culture medium	3 vials
		Gentamicin: Added to culture medium for use	1 vial, 300μL/vial
FEP Culture Bag (2L)	FP0002 - 2	In vitro culture of NK cells	3 pieces	Room Temperature

NK Large-Scale Culture Kit —— Standard Components

Product Name	Catalog Number	Application	Specification	Storage
Immune Cell Repair Medium (Phenol Red - Free)	NC0111	For pre - activation of NK cells	1 bottle, 200mL/bottle	2 ~ 8°C
NK Cell Serum - Free Medium	NC0102	In vitro culture of NK cells	2 bottles, 1L/bottle	2 ~ 8°C
NK Cell Induction and Expansion Kit (High - Performance Version) - 1L System	AN0104 - 1	YC00A: For coating culture bottles during initial culture	1 vial, 250μL/vial	-20°C
		YC00B: For addition during initial culture	1 vial, 250μL/vial
		YC00C: Addition to activation medium	1 vial, 100μL/vial
		YC005: Addition to expansion medium	1 vial
		Gentamicin: For addition to culture medium	Gentamicin: 1 vial, 300μL/vial
NK Factor YC005	YC005 - 1	Addition to expansion medium	1 vial	2 ~ 8°C
FEP Culture Bag (2L)	FP0002 - 2	In vitro culture of NK cells	1 piece	Room temperature

Usage Guidelines

The kit's modularity allows for precise control over the culture process. Begin with the Standard Components for approximately 80g of umbilical blood or 50mL of pure umbilical blood. For every additional 20g of umbilical blood beyond this, integrate an Expansion Component to ensure optimal nutrient and factor availability, enabling proportionate cell growth and achieving your target cell counts.

Q1: What samples are suitable for large-scale umbilical cord blood NK cell culture?

Samples transported within 8 hours with anticoagulant ≤28% and samples transported 8–24 hours with anticoagulant ≤25% can be cultured using autologous plasma.
For samples with incompatible anticoagulant ratios, validated commercial blood substitutes or human AB serum can replace autologous plasma.

Q2: How to assess the quality of umbilical cord blood mononuclear cells (MNCs)?

Samples with shorter transport times and lower anticoagulant ratios yield higher-quality MNCs.
After red blood cell lysis, the red blood cell ratio should be <50%.
For cryopreserved MNCs: Repair cells in immunorepair medium before culture. Post-repair spontaneous apoptosis should be ~20%.

Q3: You recommend seeding 6E7 cells/T75 flask. Can I increase or decrease the inoculum?

Not recommended. Adjusting the inoculum may cause mismatched factor quantities in the kit, compromising final cell yield or purity.

Q4: If the number of isolated MNCs is insufficient for two T75 flasks, what should I do?

Seed 1 T75 flask (6E7 cells) and supplement with 1–2 T25 flasks to utilize the remaining cells.

noculation Scheme	Inoculation Container	Inoculation per Bottle	Number of Inoculated Bottles	Required Number of Cells
Scheme 1	T75	6.00E+07	2 bottles	1.20E+08
Scheme 2	T75	6.00E+07	1 bottle	1.00E+08
Scheme 2	T25	2.00E+07	2 bottles	1.00E+08
Scheme 3	T75	6.00E+07	1 bottle	8.00E+07
Scheme 3	T25	2.00E+07	1 bottle	8.00E+07

Q5: Can cryopreserved mononuclear cells be used for large-scale umbilical cord blood NK cell culture?

Yes. Preferably use Youkang single-cell cryopreservation medium or other validated immunocyte cryopreservation media. Ensure post-thaw cell viability ≥90%. Due to cryopreservation-induced damage, a portion of cells may undergo apoptosis after inoculation. We recommend "repair before culture": Incubate thawed cells in immunorepair medium to restore viability before proceeding.

Q6: Must I strictly follow the manual’s feeding protocol?

Days 3 and 5: Follow the manual strictly for feeding and plasma supplementation.
After Day 7: Adjust feeding based on cell status (e.g., reduce volume if metabolic activity slows).

Q7: How to determine whether to continue feeding to 8L or 12L after reaching 6L?

Most qualified samples can reach 6L; some may scale to 12L. Key indicator: If cell numbers double within 2–3 days post-feeding, the culture retains strong expansion capacity—proceed with further feeding.