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HiXpan ® NK Serum-Free Culture Kit (Trophoblast Method)
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Industry Background of NK Cells
Immunotherapy is currently one of the most promising approaches for cancer treatment, with natural killer (NK) cells emerging as particularly promising effector cells. Immune cells are broadly categorized into adaptive and innate components. Innovative therapeutics targeting adaptive immune cells include cytotoxic T lymphocytes (CTLs), tumor-infiltrating lymphocytes (TILs), chimeric antigen receptor T cells (CAR-T cells), and T-cell receptors engineered T cells (TCR-T cells). Among these, CAR-T therapy is currently the most actively pursued modality; however, following CAR-T cell infusion, patients may experience two major severe adverse effects: cytokine release syndrome and neurotoxicity. Although CAR-T therapy achieves high rates of complete remission, it is associated with immune cell exhaustion, which can lead to disease relapse, and its efficacy against solid tumors remains limited due to off-target effects.
Nonspecific immune cells are primarily represented by natural killer (NK) cells. NKG2D and CD244 enhance the cytotoxic activity and cytokine production of these cells, thereby improving their antitumor efficacy. NK cells can exert their cytotoxic activity without prior stimulation, resulting in a faster response. NK cells typically do not mediate graft-versus-host disease (GVHD), making them safer. Moreover, NK cells are readily obtainable from diverse sources, including peripheral blood, umbilical cord blood, human embryonic stem cells, induced pluripotent stem cells, and even the NK-92 cell line, which makes them one of the preferred options for off-the-shelf immune cell therapies. CAR-NK cells also have limitations; for instance, the issue of NK cell immunological memory remains亟待解决 (urgently needs to be addressed). To maintain sufficient efficacy of CAR-NK cells during treatment, multiple infusions are required, and a sufficient number of high-purity, highly active effector cells must be available.
Mechanism of NK Cell Expansion and Culture
There are four major requirements for the in vitro preparation of NK cells: cell expansion quantity, NK cell purity, expression of activating receptors, and NK cell cytotoxicity. How to obtain a sufficient number of high-quality effector NK cells is one of the most critical challenges. In vitro culture of primary NK cells is broadly divided into Cytokine method and trophoblast method The primary advantage of the cytokine-based approach is its relatively well-defined composition, which facilitates regulatory oversight; however, it poses challenges for achieving rapid expansion of NK cells.
Research shows that, The trophoblast-based method enables rapid expansion of NK cells, yielding high-purity, high-quality NK cells with excellent stability, high purity, and fast expansion. Trophoblast cells must be irradiated prior to use; their safety primarily depends on whether they exhibit tumorigenic and oncogenic potential. As for residual safety concerns, if the trophoblast cells themselves can be demonstrated to be safe, then, in a certain sense, any residual cellular debris would also be considered safe. Oncogenicity refers to the ability of the “cellular components” of the test cells to induce tumor formation in host animals after inoculation; prior to inoculation, the test cells must be lysed to determine whether these cellular components possess the capacity to trigger tumorigenesis in the host. Standardized operational procedures, stringent quality control over technical manufacturing processes and storage conditions, as well as rigorous testing for mycoplasma and endotoxins, are all critical factors.
Currently, the commonly used feeder cell lines for expanding trophoblast cells via NK cells are: RPMI8866, Epstein-Barr lymphoblastoid cell line (EBV-LCL), and K562 . The feeder cells are a non-proliferating cell layer that provides extracellular secretions to support the proliferation of another cell. It is known that irradiated K562 cells sensitive to NK cells are stimulated. ， It can enhance the in vitro cytotoxic and proliferative activities of NK cells. In addition, genetic modification of K562 cells to stably express multiple cytokines, under the synergistic action of these cytokines, facilitates the targeted activation and expansion of natural killer cells derived from peripheral blood mononuclear cells, thereby significantly increasing the purity of NK cells.
IL-15 NK Cell Expansion Reagent (Trophic Cells) is a product developed by the R&D team at Heyousheng Biotechnology Co., Ltd., designed for the expansion and culture of human natural killer (NK) cells, and can Efficient expansion of NK cells derived from human peripheral blood and umbilical cord blood (NK cells can be expanded 300–400-fold through culture). ， Calculate the final harvested NK cells relative to the initial PBMCs. ) , High purity of NK cells (CD3− CD56+ expression rate can exceed 90%) 。
An independently developed, domestically produced original research system;
Suitable for the culture of NK cells from multiple sources;
After 13 days of culture, NK cell expansion reaches 300–400-fold.
The purity of expanded NK cells can reach as high as 90%;
Compatible with mainstream basal immune cell culture media (which do not contain interleukin-2); prior to use, an appropriate concentration of IL-2 must be added to the medium.
IL-15 NK Cell Expansion Reagent (Trophoblast Cells)
& Donor pre-screening
NK cells expanded under these conditions exhibit the following characteristics:
1) High amplification efficiency;
2) The effector cells exhibit high purity, reaching over 90%;
3) Simple operation; no need to add activation or amplification reagents a second time.
4) Combining with He YouSheng’s pre-screening and cryopreservation of PBMCs can deliver more stable and reproducible expansion outcomes.
Product Code Product Name Product Specifications NK881-F NK Cell Expansion Reagent (Trophoblast Cell Method, IL-15) 1 mL NK881-21-F NK Cell Expansion Reagent (Trophoblast Cell Method, IL-21) 1 mL NK881-2L IL-15 NK Cell Expansion Kit, 2 L (Trophoblast Cell Method) 1Kit NK881-21-2L IL-21 NK Cell Expansion Kit, 2 L (Trophoblast Cell Method) 1Kit No second time needed.
Add activation amplification reagent
More convenient and efficient ！
Comparison of NK Cell Expansion from Different Donors (Fresh)
Data on the expansion of cryopreserved PBMCs after thawing
Comparison of data before and after overnight culture following cryopreservation and thawing of expanded NK cells
For more details on HeYouSheng Bio’s IL-15 NK cell expansion reagent (nourishing cells), please contact our Head of Business Development, or scan the QR code below to request a free trial sample.
Contact: Mr. Xie, 15201775322.
Welcome to apply for a trial!
Download related documents for NK cell expansion reagents (trophoblast cell method):
Instruction Manual for IL-21 (2L) NK Cell Expansion Reagent Culture Kit (Support Cells)
Instruction Manual for IL-15 (2L) NK Cell Expansion Reagent Culture Kit (Stromal Cells)
IL-21 NK Cell Expansion Feeder (Stromal Cells) Instruction Manual
IL-15 NK Cell Expansion Feeder (Trophoblast Cells) Instruction Manual
IL-15 NK Cell Expansion Kit (Trophoblast Cells), 1 L – Instruction Manual
IL-21 NK Cell Expansion Kit (Trophoblast Cells), 1 L – Instruction Manual
NK Cell Expansion Kit (Trophoblast Cell Method) – Expansion Tips
Related document downloads:
Instruction Manual for IL-21 (2L) NK Cell Expansion Reagent Culture Kit (Support Cells)
Instruction Manual for IL-15 (2L) NK Cell Expansion Reagent Culture Kit (Stromal Cells)
IL-21 NK Cell Expansion Feeder (Trophoblast Cells) Instruction Manual
IL-15 NK Cell Expansion Feeder (Trophoblast Cells) Instruction Manual
IL-15 NK Cell Expansion Kit (Trophoblast Cells), 1 L – Instruction Manual
IL-21 NK Cell Expansion Kit (Trophoblast Cells), 1 L – Instruction Manual
NK Cell Expansion Kit (Trophoblast Cell Method) – Expansion Tips
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