Efficient Expansion of Human Umbilical Cord Blood- Derived NK Cells Ex Vivo without Requiring Feeder Layers
Journal Title: Biomedical Journal of Scientific & Technical Research (BJSTR) - Year 2018, Vol 8, Issue 5
Abstract
Natural killer (NK) cells are central components of the innate immunity. They have ability to kill a wide range of cancer cells and are a promising tool for both autologous and allogeneic immune enhancement therapy in cancer treatment. Actually, NK cells can be derived from multiple sources such as: peripheral blood, cord blood... Among these, cord blood (CB) is known as an ideal for NK cells expansion because it occupies a higher percentage of NK cells compare to the peripheral blood and is a rich source of hematopoietic stem cell as well as progenitor immune cells. The objectives of this study were to invent a reasonable approach for expanding a relevant number of NK cells from human cord blood to clinical application. Methods: At the initial step, cord blood was collected from the patients and then the mononuclear cells were obtained by density centrifugation with Ficoll. After that, these cells were cultured in 2 stages: stimulation and expansion with inactivated autologous plasma and BINKIT expansion kit contains several kinds of growth factors which are specified for NK cells. The immunophenotype of NK cells was analyzed every 2 days through %CD56+CD3- by Flow cytometry technique. The ex-vivo activation and expansion of NK cells was performed in GMP grade -clean room for about 3 weeks. Results: After culturing periods, the average NK cell count post-expansion was 2040.6 ± 1463.5 x10A6 and the average cell fold expansion was 814.4 ± 560.9, the expanded cells presented 90.6% ± 8.9% purify of CD56+CD3-, the percentage of CD56 bright CD3- cells was 84.2% ± 14.4%. Conclusion: Human cord blood has a significant potential to expand NK cells to relevant number for clinical use and our method are suitable for getting a relevant number and quality of NK cells to the clinical use.Nowadays, cancer is still a leading cause of death in the world. However, the standard treatments for cancer such as surgical therapy, radiotherapy and chemotherapy usually bring multiple side-effects to patients. In some recent years, the invention of cell therapy has opened brighter future in some life-threatening diseases treatment, especially cancer. This approach helps patients to avoid immunodeficiency through sufficient provision of immune cells as well as stimulating these cells to kill specifically cancer cells. Clinical trials of cell therapy for treating many different cancers are currently ongoing. In fact, there are many types of immune cells which are used for cell therapy such as: NK cells, cytotoxic T cells (CTL), dendritic cells (DC), y5T cells and a^T cells [1]. Natural Killer (NK) cells are a subtype of type 1 ILCs (innate lymphoid cells), which play significant roles in innate immune responses. NK cells can produce cytokine and have cytotoxicity to kill both viral infected and tumour cells. Because of the ability to secrete immunostimulatory cytokines like IFN-y, NK cells can manipulate not only tumour growth but also metastasis [2]. Therefore, NK cells have been known as promising tools for cell therapies in cancer treatment. Initial clinical trials have demonstrated that the infusion of NK cells is entirely possible and safe with no side effects and minimal toxicity to patients [3]. NK cells can be determined by the expression of CD56 and the absence of the T cell marker CD3 on their surface. Besides, most human blood NK cells also express CD16, which is involved in recognition of antibody-coated cells [4]. NK cells can be divided into two main types: CD56bright and CD56dim populations [5,6]. The CD56bright cells have low or absent expression of CD16 and killer-cell immunoglobulin-like receptors (KIR), whereas the CD56dim cells express both CD16 and KIR [7,6].
Authors and Affiliations
Chu Thi Thao, Do Thi Hoai Thu, Bui Viet Anh, Truong Linh Huyen, Nguyen Van Phong, Nguyen Thanh Liem, Hoang Thi My Nhung
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