CAR-T cell therapy represents a breakthrough in cancer treatment, yet its implementation in developing countries remains challenging due to technical and infrastructural barriers. This study aimed to establish clinical-scale CAR-T production in Kazakhstan, a country with no prior experience in advanced cell and gene therapies. We implemented a complete CAR-T manufacturing pipeline, including in-house lentiviral vector (LV) production and automated CAR-T cell processing using the CliniMACS Prodigy system. Two anti-CD19 CAR LVs were used, one modeled after FDA-approved Kymriah (4-1BB costimulation) and another replicating Yescarta (CD28 costimulation). The vector produced locally achieved functional titers of 1.5 × 10¹⁰ TU/mL after concentration. Twelve clinical-scale CAR-T products were manufactured, exhibiting a memory-skewed T-cell phenotype. Functional assessments revealed that CD28-based CAR-T cells produced significantly higher Th1 cytokines (IFN-γ, TNF-α, IL-2; p < 0.05) than 4-1BB-based cells, though both demonstrated comparable cytotoxicity against CD19+ targets. These findings demonstrate the feasibility of establishing CAR-T production in resource-limited settings using a decentralized manufacturing framework. This work provides a scalable model of CAR-T therapy production in developing regions, suitable for clinical implementation using the hospital exemption framework. Critical gaps in access to advanced immunotherapies, including CAR-T, in the Central Eurasia region are addressed.

Background/Objectives: Non-receptive endometrium is associated with recurrent implantation failure, which leads to a decrease in the frequency of pregnancy during IVF; therefore, new treatment methods such as the use of Platelet-Rich Plasma (PRP) are gaining popularity in the treatment of infertility in women with repeated unsuccessful IVF attempts. Methods: A total of 38 women were included in this study, with the main complaint being the inability to conceive or maintain pregnancy. Medical examination, laboratory tests, ultrasound of the pelvic organs and hysteroscopy were performed. After that, whole blood was taken to prepare an autologous PRP; then, the PRP was inserted into the uterine using an intrauterine catheter in the first phase of the menstrual cycle (1–7 procedures). The primary outcome of this study was an increase in endometrial thickness and improvement of the receptive endometrial layer. The secondary outcome was pregnancy rate. This was a single-center prospective interventional clinical study. Results: Statistical analysis of changes in endometrial thickness after PRP therapy showed that endometrial thickness indicators after treatment significantly exceeded the values before the intervention. This may be evidence of the effectiveness of PRP therapy for thin endometrium. When analyzing pregnancy status, it was noted that after receiving PRP, more than half (56% of cases) became pregnant and the majority of them successfully gave birth. Conclusions: Based on the results of our study, we can conclude that intrauterine injection of PRP may be a new therapeutic approach in the treatment of thin endometrium and associated infertility. The use of PRP demonstrated effectiveness in increasing the thickness of the endometrium, regardless of pregnancy, while the secondary indicator was the frequency of successful pregnancies among the participants.

Introduction. Automated leukapheresis is the initial step in the production of CAR-T cell products and provides a concentrated fraction of mononuclear cells used for subsequent isolation, genetic modification, and expansion of T-lymphocytes. Despite the widespread introduction of CAR-T therapy into clinical practice, the questions of standardization of the leukapheresis procedure and assessment of the quality of the initial cellular material are still insufficiently investigated, especially in the organization of academic CAR-T cell production. Objective. To evaluate the quality of the mononuclear cell concentrate obtained by automated leukapheresis and determine the possibility of its use as the initial material for CAR-T cell production within the implementation of the national scientific and technical program of the Republic of Kazakhstan. Materials and methods. A pilot study was conducted at the Scientific and Production Center for Transfusiology. The study included 12 participants: 7 healthy donors and 5 patients with oncohematological diseases. Automated leukapheresis was performed using the Spectra Optia® system (Terumo BCT, USA) with the Mononuclear Cell Collection program. The technical success of the procedure, main technological parameters, cellular composition of the obtained product, number of viable CD45⁺/CD3⁺ T lymphocytes, and results of microbiological control were evaluated. Results. All automated leukapheresis procedures were completed in full without technical complications. The obtained mononuclear cell concentrates were suitable for subsequent CAR-T cell production. The mean volume of processed whole blood was 9549.5 mL, and the mean duration of the procedure was 230.8 min. The number of leukocytes in the obtained products ranged from 6.5×10⁹ to 32.1×10⁹ cells per dose, with a mean of 14.4±8.0×10⁹. The CD3⁺ T-lymphocyte content ranged from 129.1×10⁷ to 1629.7×10⁷ cells per dose, with a mean of 607.2±526.6×10⁷. Sterility of the cellular product was confirmed in all cases. Conclusion. The obtained results confirm the possibility of obtaining a mononuclear cell concentrate by automated leukapheresis that meets the requirements for subsequent CAR-T cell production. The developed algorithm for performing the procedure, laboratory quality assessment, and transfer of the cellular product can be used for further standardization of the collection of initial cellular material and development of a national platform for academic CAR-T cell production. Keywords: CAR-T cell therapy, automated leukapheresis, mononuclear cell concentrate, CD3⁺ T lymphocytes, cell therapy.

Introduction: CAR-T cell therapy is the newest and most effective treatment method for resistant forms of B-cell hematological malignancies; however, it has until now remained unavailable in Kazakhstan. The first and critically important step in the manufacturing of CAR-T products is apheresis-based leukapheresis — a procedure that determines the quality of the starting cellular material for genetic modification of T lymphocytes. In the Republic of Kazakhstan, this procedure was performed within the framework of a national scientific and technical program for the implementation of CAR-T therapy. Objective: To study the parameters of apheresis-based leukapheresis and to characterize the cellular composition of the obtained mononuclear cell concentrate in order to assess its suitability as starting material for the manufacturing of CAR-T products. Materials and Methods: At the Department of Cell Technologies of the Scientific and Production Center of Transfusiology (Astana), 12 leukapheresis procedures were performed in 7 healthy donors and 5 patients with oncohematological diseases. An automated separator Spectra Optia (Terumo BCT, USA) was used with the Mononuclear Cell Collection program. The volume of processed blood, procedure duration, product cellularity, CD3⁺ T-cell content, and sterility were assessed. Results: All 12 procedures were completed without complications. The mean volume of processed blood was 9549.5 mL (1.0–2.6 blood volumes), and the mean duration was 230.8 min. Cell yield varied: total leukocyte count ranged from 6.5×10⁹ to 32.1×10⁹ (mean 14.4±8.0×10⁹), and CD3⁺ T-lymphocyte content ranged from 129.1×10⁷ to 1629.7×10⁷ cells per dose. Products from all participants were sterile and deemed suitable for CAR-T cell manufacturing. Differences were noted between the donor and patient groups; however, due to the small sample size, statistical analysis was not performed. Conclusions: Apheresis-based leukapheresis yields a cellular concentrate that fully meets the requirements for initiating the manufacturing of CAR-T products. This work represents the first standardized protocol in Kazakhstan for obtaining clinically significant cellular material for CAR-T cell manufacturing and lays the foundation for the development of national quality criteria. Funding: Program-Targeted Funding of the Ministry of Health of the Republic of Kazakhstan BR25293293 "Implementation of chimeric antigen receptor (CAR)-T cell therapy technology for hematological tumors into practical healthcare."