Cell Bank Research Articles

Stem cell therapy: Advances and future directions in regenerative medicine: A review

Stem cell treatment, an important part of regenerative medicine that may change the way many illnesses and injuries are treated in the future. This review article gives an in-depth analysis of the many kinds of stem cells, including adult, induced pluripotent, and embryonic stem cells, as well as their background, properties, and potential for differentiation. Successful hematopoietic stem cell transplantation, tissue engineering, medications for neurological illnesses, and management of autoimmune diseases are only a few of the current applications of stem cells in therapy that we explore. The report also discusses cutting-edge techniques in stem cell research, such as 3D bioprinting, the game-changing CRISPR-Cas9 gene-editing approach, and the manipulation of stem cells using tiny chemicals. However, there are several obstacles on the road to stem cell treatment. We explain how ethical concerns, immunological rejection, and tumorigenicity have influenced public opinion. Future developments in stem cell therapy will be discussed, including the role of stem cell banking, tissue engineering, personalized medicine, and artificial intelligence. In conclusion, despite substantial progress, barriers still need to be eliminated before stem cell treatment can realize its promise to fundamentally alter regenerative medicine.

A cell bank paradigm for preclinical evaluation of an analogous cellular product for an allogeneic cell therapy

Toward the translation of allogeneic cell therapy products, cell banks are needed not only to manufacture the final human product but also during the preclinical evaluation of an animal-based analogous cellular product (ACP). These cell banks need to be established at both the master cell bank (MCB) level and the working cell bank (WCB) level. Inasmuch as most of the development of cell therapy products is at academic centers, it is imperative that academic researchers understand how to establish MCBs and WCBs within an academic environment. To illustrate this process, using articular cartilage as the model, a cell bank for an ACP was developed (MCBs at passage 2, WCBs at passage 5) to produce self-assembled neocartilage for preclinical evaluation (constructs at passage 7). The cell bank system is estimated to be able to produce between 160 000 and 400 000 constructs for each of the six MCBs. Overall, the ACP cell bank yielded constructs that are analogous to the intended human product, which is critical toward conducting preclinical evaluations of the ACP for inclusion in an Investigational New Drug application to the FDA.

Advantages of pooling of human bone marrow-derived mesenchymal stromal cells from different donors versus single-donor MSCs

Mesenchymal stromal cells (MSC) from adult bone marrow are the most commonly used cells in clinical trials. MSCs from single donors are the preferred starting material but suffer from a major setback of being heterogeneous that results in unpredictable and inconsistent clinical outcomes. To overcome this, we developed a method of pooling MSCs from different donors and created cell banks to cater clinical needs. Initially, the master cell banks (MCBs) were created at passage 1 (P1) from the bone marrow MSCs isolated from of nine different donors. At this stage, MCBs from three different donors were mixed in equal proportion and expanded till P3 to create working cell banks. Further, the pooled cells and individual donor MSCs were expanded till P5 and cryopreserved and extensively characterised. There was a large heterogeneity among the individual donor MSCs in terms of growth kinetics (90% Coefficient of variation (CV) for cell yield and 44% CV for population doubling time at P5), immunosuppressive ability (30% CV at 1:1 and 300% CV at 1:10 ratio), and the angiogenic factor secretion potential (20% CV for VEGF and71% CV for SDF-1). Comparatively, the pooled cells have more stable profiles (60% CV for cell yield and 7% CV for population doubling time at P5) and exhibit better immunosuppressive ability (15% CV at 1:1 and 32% CV at 1:10 ratio ) and consistent secretion of angiogenic factors (16% CV for VEGF and 51% CV for SDF-1). Further pooling does not compromise the trilineage differentiation capacity or phenotypic marker expression of the MSCs. The senescence and in vitro tumourigenicity characteristics of the pooled cells are also similar to those of individual donor MSCs. We conclude that pooling of MSCs from three different donors reduces heterogeneity among individual donors and produces MSCs with a consistent secretion and higher immunosuppressive profile.

Knowledge and attitude on umbilical cord blood stem cell therapy among bachelor nursing students in Lalitpur Metropolitan City

Background: Stem cells are immature cells that have ability to mature into different types of functioning cells as needed by body. Umbilical cord blood is one of the richest sources of stem cells. It is vital for nurses to keep up-to-date of current advances in stem cell science and nurses can play crucial role in providing first-hand information to public about stem cell and umbilical cord blood banking. Therefore, this study is aimed to identify the knowledge and attitude regarding umbilical cord blood stem cell therapy among bachelor nursing students. Method: A descriptive cross sectional research design was used to assess the knowledge and attitude of 263 nurses studying bachelor in nursing science and post basic bachelor in nursing in different selected nursing college of Lalitpur metropolitan city. Multi stage cluster sampling was used to select respondents. Data was collected using self-developed self-administered google form questionnaire. Results: Most of the nurses had poor knowledge (49.4%) and few nurses had good knowledge (6.8%) however majority of nurses (93.9%) had favourable attitude regarding umbilical cord blood stem cell therapy. Regarding association between variable, there is no any association between knowledge level and attitude (p=0.224). Conclusion: Nurses had poor knowledge and favourable attitude regarding umbilical cord blood stem cell therapy. Keywords: Nurses,Stem cells, Umbilical cord blood

Significance of Dental Stem Cells in Dentistry and Stem Cell Banking

Dentists play a crucial role in collecting stem cells for dental treatments and the potential treatment of medical diseases. Considering dental stem cells (DSCs) in terms of their increasing potential in medicine, adequate knowledge and positive attitudes of dentists toward DSCs and their use in regenerative therapies are extremely important. The immense potential of DSCs in regenerative medicine applications for various treatments suggests their significant role in both dental treatments and innovative approaches to treating various diseases. “Dental banking” emerged as a result of DSCs’ use in medical applications and their numerous clinical advantages across regenerative medical domains. DSC banking is a service that makes it possible to store and utilize DSCs for potential medical applications. However, further clinical research is needed to advance the applications of DSCs in treatment. This review aims to summarize the current literature on dental stem cell banking applications and usage areas of DSCs in dentistry.

A phase I/II study of adoptive SARS-CoV-2-specific T cells in immunocompromised hosts with or at risk of severe COVID-19 infection

BackgroundPost-transplant or hematological cancer patients have a higher risk of mortality after infection with ancestral and early variants of severe acute respiratory syndrome (SARS)-CoV-2. Adoptive cell therapy (ACT) with virus-specific T cells (VSTs) could augment endogenous T cell immunity to avoid disease deterioration before viral clearance. MethodsWe established a third-party SARS-CoV-2-specific T cell (COVID-T) bank in 2020 (NCT04351659) using convalescent and/or vaccinated donors. In a phase I/II study (NCT04457726), 13 adult and pediatric patients, acutely positive for SARS-CoV-2 and predicted to have a high chance of mortality, were recruited from September 2021 to February 2022. Twelve patients received a single dose of COVID-T cells, matched on at least 1 HLA. ResultsA dose of either 75,000 or 150,000 IFN-γ+CD3+ cells/m2 SARS-COV-2-specific T cells did not cause cytokine release syndrome, acute respiratory distress syndrome, or graft-versus-host disease. In the 8 patients who had detectable donor SARS-COV-2-specific T cells after ACT, none progressed to severe disease or died with COVID-19. In contrast, among the other four patients without evidence of donor micro-chimerism, two died of COVID-19. ConclusionsLong-acting third-party VSTs from convalescent or vaccinated donors could be expediently produced and might be clinically useful in future pandemics, particularly before global vaccination is implemented.

Study on the inhibitory and pro-apoptotic effects of different concentrations of total tanshinone alone and in combination with tyrosine kinase inhibitors on human myeloid leukemia cell lines

Objective: To explore the effect and investigate the molecular mechanism of different concentrations of total tanshinones alone and in combination with tyrosine kinase inhibitors (TKIs) on the proliferation inhibition and apoptosis of human myeloid leukemia cell lines. Methods: K562 and Kasumi-1 cell lines were purchased from the Shanghai Cell Bank of the Chinese Academy of Sciences, and the TKIs-resistant strain K562/T315I cell line was constructed in Molecular Medicine Research Center, Beijing Lu Daopei Institute of Hematology. Logarithmic growth phase cells were taken and divided into intervention groups with total tanshinone of 0, 2.19, 4.38, 8.75, 17.50 and 35.00 μg/ml intervention groups, which were inoculated in 96-well plates at a density of 1×104 cells/well and exposed to the drug for 24 h, and a control group treated with dimethyl sulfoxide was also set up simultaneously. All experiments were repeated independently 3-5 times. The proliferative activity of the cells was assessed using the CCK-8 assay, the apoptotic rates were measured by flow cytometry, and the expression levels of apoptosis-regulating proteins Bcl-2 and Bax were analyzed by Western blotting. The cell lines treated and untreated with total tanshinone were subjected to transcriptome sequencing and gene set enrichment analysis to identify differentially expressed genes. Results: The half-inhibitory concentration (IC50) values of 8.75 μg/ml total tanshinone at 24 h for K562, K562/T315I and Kasumi-1 cells were (4.11±0.02), (4.95±0.04) and (3.98±0.01) μg/ml, respectively. When combined with 0.25 μmol/L imatinib, 8.75 μg/ml total tanshinone could enhance the induction of apoptosis effects on K562 and K562/T315I cell lines. After being treated with 4.38, 8.75, and 17.50 μg/ml of total tanshinone for 24 h, compared with the control group, total tanshinone upregulated the expression level of Bax protein, downregulated the expression level of Bcl-2 protein, and decreased the Bcl-2/Bax ratio (all P<0.05). Total tanshinone inhibited the proliferation-related signaling pathway and DNA damage repair pathway of myeloid leukemia cell lines, and activated the signaling pathway that induces apoptosis in leukemia cells. Conclusion: Different concentrations of total tanshinoneinhibites proliferation and promote apoptosis in K562, Kasumi-1 and TKIs-resistant K562/T315I cell lines, and further enhance the anti-leukemic effect when combined with TKIs.

Cytotoxic effect of silver nanoparticles biosynthesized from Hirudo medicinalis saliva on HepG2 cells

Primary hepatocellular carcinoma is a devastating type of liver cancer. Silver nanoparticles (AgNPs) have been assessed for a variety of purposes, including being tested as an anticancer agent. The aim of this study was to assess the cytotoxicity of AgNPs that were biosynthesized from leech saliva on HepG2 cells, through the undertaking of a simple MTT assay. HepG2 cells were obtained from the cell bank of the Pasteur Institute of Iran. In this study, AgNP-treated HepG2 cells were cultured at a density of 104 cells per well, and 100 μL of MTT at a concentration of 0.5 mg/mL were added to each well; the treated cells were then let to incubate for 4 h. Subsequently, a plate reader device operating at a wavelength of 570 nm was used in order to determine the concentration of the chemical dissolved in isopropanol. Representative images of the cells show remarkable changes in their morphology at AgNP concentrations of 25 and 50 μg/mL. At 48 h, the nanoparticle’s IC50 value was 50 μg/mL. Our study shows that leech salivary extract-derived AgNPs are cytotoxic to HepG2 cells.

Validation of a next generation sequencing method for adventitious virus detection: Demonstration of sensitivity in multiple cell lines

In the past few years NGS has become the technology of choice to replace animal-based virus safety methods and this has been strengthened by the recent revision to the ICHQ5A virus safety chapter. Here we describe the validation of an NGS method using an agnostic analysis to detect and identify RNA virus and actively replicating DNA virus contaminants in cell banks. We report the results of the validation of each step in the sequencing process that established quality criteria to ensure consistent sequencing data. Furthermore, the validation of the analysis algorithm designed to identify virus specific sequences is described along with steps undertaken to ensure the integrity of the sequencing data from generation to analysis. Lastly, the validated sequencing and analysis systems were used to establish a limit of detection (LOD) for model viruses in cells that are commonly used in biomanufacturing. The LOD from these studies ranged from 1E+03 to 1E+04 genome copies and were dependent on the virus type with little variability between the different cell types. Thus, the validation of the NGS method for adventitious agent testing and the establishment of a general LOD for cell-based samples provides a suitable alternative to traditional virus detection methods.

Generation and characterization of GATA6-specific EGFP expressing human induced pluripotent stem cell line, KSCBi017-A-1, using CRISPR/Cas9

GATA6 is expressed during early embryogenesis and localizes to endoderm- and mesoderm-derived tissues during later embryogenesis. Here, we established a human induced pluripotent stem cell (hiPSC) line expressing EGFP under GATA6 gene. EGFP coding sequence was introduced into the C-terminus of GATA6 in KSCBi017-A hiPSCs through homologous recombination using CRISPR/Cas9 system. The successfully edited line, KSCBi017-A-1, was selected and confirmed by sequencing. The line had a normal karyotype and exhibited potential to differentiate into three germ layers while it expressed EGFP upon endoderm induction. KSCBi017-A-1 cells can be used to monitor the expression of GATA6 during differentiation. This cell line is available from Korea National Stem Cell Bank.

Genetic diversity among the present Japanese population: evidence from genotyping of human cell lines established in Japan

Japan is often assumed to have a highly homogeneous ethnic population, because it is an island country. This is evident in human cell lines collected from cell banks; however, these genotypes have not been thoroughly characterized. To examine the population genotypes of human cell lines established in Japan, we conducted SNP genotyping on 57 noncancerous cell lines and 43 lung cancer cell lines. Analysis of biogeographic ancestry revealed that 58 cell lines had non-admixed Japanese genotypes, 21 cell lines had an admixture of Japanese and East Asian genotypes, and the remaining 21 cell lines had East Asian genotypes. The proportion of non-admixed Japanese genotypes was similar between lung cancer and noncancerous cell lines, suggesting that patients in Japan may not exclusively have Japanese genotypes. This could influence the incidence of inherited diseases and should be taken into account in personalized medicine tailored to genetic background. The genetic makeup of the present-day Japanese population cannot be fully explained by the ancestral Jomon and Yayoi lineages. Instead, it is necessary to consider a certain level of genetic admixture between Japanese and neighboring Asian populations. Our study revealed genetic variation among human cell lines derived from Japanese individuals, reflecting the diversity present within the Japanese population.

Exploring Parental Knowledge, Attitudes, and Factors Influencing Decision-Making in Stem Cell Banking: Rising the Future of Medical Treatment.

Stem cellbanking (SCB) is a promising area of modern medicine with the potential to yield innovative treatments and cures. To effectively educate parents and implement laws and regulations that address parental concerns and encourage informed decision-making, it is imperative to emphasize parental viewpoints and their consequences for future healthcare. The study aims to establish the Saudi Arabian population's level of understanding regarding SCB and to comprehend the elements influencing parental knowledge, attitudes, and SCB decision-making processes. A cross-sectional study was conducted among the population in the Makkah region of Saudi Arabia. Demographic data, knowledge levels, attitudes, and decision-making variables were gathered from 380 respondents. The study reveals a lack in their comprehension of the objectives and possible uses of SCB, together with the main sources of information on those banks and conveniently available banking choices. It showed varied results regarding attitudes about considering an SCB for their children. In addition, the majority of respondents had not made a consent decision about SCB for their children. It also illuminates the factors that could influence participants' decisions about SCBfor their children and shows that a lack of information and understanding is the main obstacle faced by parents regarding SCB. It highlights that participants were generally in favor of learning more about SCB for their children. This study broadens our understanding of parental decision-making toward SCB and clarifies the elements influencing parents' opinions and worries and offers significant ramifications for lawmakers, medical professionals, and SCB. These implications can be utilized to enhance communication strategies, create instructional programs, and ease the fears of concerned parents.

The Ethical Challenges of Umbilical Cord Blood Stem Cell Banking: A Qualitative Study

Background: Umbilical cord blood serves as a potent source of hematopoietic stem cells, utilized in treating specific diseases and preserved in blood banks. Objectives: This study aims to explore the ethical issues associated with these repositories. Methods: This qualitative research involved conducting 14 interviews with experts and the families of recipients or donors of umbilical cord blood. Participants were selected purposefully from May to November 2021. Results: The investigation identified five principal ethical challenges: presentation of information, interpersonal relationships among involved parties, respect for human dignity, adherence to rules and regulations, and the handling of tissue samples. These challenges encompassed various categories and subcategories, including informed consent, communication and advertising of information, financial considerations, conflicts of interest, professional interactions, ownership rights concerning the child and parents, privacy and confidentiality issues, compliance with regulations and guidelines, and the clinical and research uses of the tissue samples. Conclusions: The study unveiled significant ethical concerns in the domain of umbilical cord blood stem cell banking. Addressing these ethical dilemmas necessitates the involvement of health policymakers and medical ethics experts, along with a comprehensive understanding of these banks' multifaceted nature by the community.

Establishment of Big Bone chicken fibroblast cell bank and study of its biological characteristics

A fibroblast bank of Big Bone chicken was established using tissue adherent culture method. This cell bank included 29 embryo samples, had stocks of 147 cryogenically-preserved vials each containing 2~3Å~106 cells, and met all the cell line quality control standards established by the American Type Culture Collection (ATCC). The cells cultured in vitro showed the typical morphology of fibroblasts. The growth curve assumed the “S” shape, and the cell population doubling time (PDT) was about 30h. Cell viability was 98.1% before cryopreservation and 96.6% after recovery. All tests for microbial contamination were negative. The isoenzyme pattern was of species specificity. The frequency of diploid cells was 91%. The transfection efficiency of three fluorescent protein genes fluctuated between 10.6% and 26.5%. These results showed that the cells cultured in vitro grew well and had stable genetic properties. This research thus does not only preserve the poultry genetic resources of Big Bone chicken at the cell level, but also opens new ways for preserving important genetic resources of endangered animals in the form of somatic cells.

Establishment and characterization of NCC-ASPS2-C1: a novel patient-derived cell line of alveolar soft part sarcoma.

Alveolar soft part sarcoma (ASPS) is a rare mesenchymal tumor characterized by rearrangement of the ASPSCR1 and TFE3 genes and a histologically distinctive pseudoalveolar pattern. ASPS progresses slowly, but is prone to late metastasis. As ASPS is refractory to conventional chemotherapy, the only curative treatment is complete surgical resection. The prognosis of advanced and metastatic cases is poor, highlighting the need for preclinical research to develop appropriate treatment options. However, ASPS is extremely rare, accounting for < 1% of all soft tissue sarcomas, and only one patient-derived ASPS cell line is available from public cell banks worldwide for research. This study reports the establishment of a novel ASPS cell line derived from the primary tumor tissue of an ASPS patient, named NCC-ASPS2-C1. This cell line retains the ASPSCR1-TFE3 fusion gene, which is characteristic of ASPS. The characterization of this cell line revealed stable growth, spheroid formation, and invasive properties. By screening a drug library using NCC-ASPS2-C1, we identified several drugs that inhibited the proliferation of ASPS cells. In conclusion, the establishment of NCC-ASPS2-C1 provides a valuable resource for advancing ASPS research and developing novel treatments for this challenging disease.

Establishment and characterization of two novel patient-derived cell lines from giant cell tumor of bone: NCC-GCTB8-C1 and NCC-GCTB9-C1.

Giant cell tumor of bone (GCTB) is a rare osteolytic bone tumor consisting of mononuclear stromal cells, macrophages, and osteoclast-like giant cells. Although GCTB predominantly exhibits benign behavior, the tumor carries a significant risk of high local recurrence. Furthermore, GCTB can occasionally undergo malignant transformation and distal metastasis, making it potentially fatal. The standard treatment is complete surgical resection; nonetheless, an optimal treatment strategy for advanced GCTB remains unestablished, necessitating expanded preclinical research to identify appropriate therapeutic options. However, only one GCTB cell line is publicly available from a cell bank for research use worldwide. The present study reports the establishment of two novel cell lines, NCC-GCTB8-C1 and NCC-GCTB9-C1, derived from the primary tumor tissues of two patients with GCTB. Both cell lines maintained the hallmark mutation in the H3-3A gene, which is associated with tumor formation and development in GCTB. Characterization of these cell lines revealed their steady growth, spheroid-formation capability, and invasive traits. Potential therapeutic agents were identified via extensive drug screening of the two cell lines and seven previously established GCTB cell lines. Among the 214 antitumor agents tested, romidepsin, a histone deacetylase inhibitor, and mitoxantrone, a topoisomerase inhibitor, were identified as potential therapeutic agents against GCTB. Conclusively, the establishment of NCC-GCTB8-C1 and NCC-GCTB9-C1 provides novel and crucial resources that are expected to advance GCTB research and potentially revolutionize treatment strategies.

The construction of modular universal chimeric antigen receptor T (MU-CAR-T) cells by covalent linkage of allogeneic T cells and various antibody fragments

BackgroundChimeric antigen receptor-T (CAR-T) cells therapy is one of the novel immunotherapeutic approaches with significant clinical success. However, their applications are limited because of long preparation time, high cost, and interpersonal variations. Although the manufacture of universal CAR-T (U-CAR-T) cells have significantly improved, they are still not a stable and unified cell bank.MethodsHere, we tried to further improve the convenience and flexibility of U-CAR-T cells by constructing novel modular universal CAR-T (MU-CAR-T) cells. For this purpose, we initially screened healthy donors and cultured their T cells to obtain a higher proportion of stem cell-like memory T (TSCM) cells, which exhibit robust self-renewal capacity, sustainability and cytotoxicity. To reduce the alloreactivity, the T cells were further edited by double knockout of the T cell receptor (TCR) and class I human leukocyte antigen (HLA-I) genes utilizing the CRISPR/Cas9 system. The well-growing and genetically stable universal cells carrying the CAR-moiety were then stored as a stable and unified cell bank. Subsequently, the SDcatcher/GVoptiTag system, which generate an isopeptide bond, was used to covalently connect the purified scFvs of antibody targeting different antigens to the recovered CAR-T cells.ResultsThe resulting CAR-T cells can perform different functions by specifically targeting various cells, such as the eradication of human immunodeficiency virus type 1 (HIV-1)-latenly-infected cells or elimination of T lymphoma cells, with similar efficiency as the traditional CAR-T cells did.ConclusionTaken together, our strategy allows the production of CAR-T cells more modularization, and makes the quality control and pharmaceutic manufacture of CAR-T cells more feasible.

Good Manufacturing Practice–compliant human induced pluripotent stem cells: from bench to putative clinical products

Background aimsFew human induced pluripotent stem cell (hiPSC) lines are Good Manufacturing Practice (GMP)-compliant, limiting the clinical use of hiPSC-derived products. Here, we addressed this by establishing and validating an in-house platform to produce GMP-compliant hiPSCs that would be appropriate for producing both allogeneic and autologous hiPSC-derived products. MethodsOur standard research protocol for hiPSCs production was adapted and translated into a GMP-compliant platform. In addition to the generation of GMP-compliant hiPSC, the platform entails the methodology for donor recruitment, consent and screening, donor material procurement, hiPSCs manufacture, in-process control, specific QC test validation, QC testing, product release, hiPSCs storage and stability testing. For platform validation, one test run and three production runs were performed. Highest-quality lines were selected to establish master cell banks (MCBs). ResultsTwo MCBs were successfully released under GMP conditions. They demonstrated safety (sterility, negative mycoplasma, endotoxins <5.0 EU/mL and negative adventitious agents), cell identity (>75% of cells expressing markers of undifferentiated state, identical STR profile, normal karyotype in >20 metaphases), purity (negative residual vectors and no plasmid integration in the genome) and potency (expression of at least two of the three markers for each of the three germ layers). In addition, directed differentiation to somitoids (skeletal muscle precursors) and six potential clinical products from all three germ layers was achieved: pancreatic islets (endoderm), kidney organoids and cardiomyocytes (mesoderm), and keratinocytes, GABAergic interneurons and inner-ear organoids (ectoderm). ConclusionsWe successfully developed and validated a platform for generating GMP-compliant hiPSC lines. The two MCBs released were shown to differentiate into clinical products relevant for our own and other regenerative medicine interests.

Process Intensification by Inoculating Antibody Production Bioreactors Directly from Cryovials

AbstractThere are several ways to make antibody production processes more efficient and less expensive through process intensification. In this study, an ultra‐high cell density working cell bank was used to inoculate production bioreactors directly with cells from cryovials. After lab‐scale experiments, a fed‐batch pilot‐scale experiment was successfully conducted as a proof‐of‐concept. In addition, cells were cultivated in perfusion mode at 2 L scale. The intensification approach presented here allows the complete elimination of the inoculum production outside the production bioreactor and the creation of additional production capacity by saving time and space. In perfusion mode, an increase in bioreactor productivity of over 180% can be achieved compared to fed‐batch mode.

A Study to Assess the Effectiveness of Structured Teaching Programme on Level of Knowledge Regarding Menstrual Blood Stem Cell Banking among Staff Nurses Working at Selected Hospitals of Valsad District

Introduction: Menstrual blood stem cell banking allows women to store their menstrual blood under suitable conditions and preserve it for future use. Aim: This study aims to assess the effectiveness of a structured teaching program on the level of knowledge among staff nurses working at selected hospitals of the Valsad district. Methodology: Quantitative approach with a pre-experimental research design by one-group pre-test and post-test was used. A total of 100 staff nurses were selected for data collection. Self-structured questionnaires were used to assess level of knowledge and association with socio-demographic variables. After conducting a pre-test, a structured teaching program on menstrual blood stem cell banking was administered to the participants and post-test was conducted after a relapse of seven days. Results: Overall level of knowledge in Pre-test shows that 9% had average knowledge and 91% had poor knowledge and none had good knowledge. In Post-test, None of the Staff Nurse is had poor knowledge, 61% of had average knowledge and 39% had good knowledge regarding Menstrual blood stem cell banking. This present study shows the Mean of Pre-test was 6.36 and significantly improved to 17.42 in Post-test with the mean difference of 11.06. The standard deviation of Pre-test was 3.76 and Post-test was 2.4. The computed value of t= 25.04 which is greater than the table value of t= 1.645 at the level of p at 0.05. There was a significant association between the pre-test knowledge score of staff nurses and selected socio-demographic variables, including educational status and previous knowledge about menstrual blood stem cell banking by chi- square method. Conclusion: The study concluded that the structured teaching program is effective in improving the level of knowledge of staff nurses regarding menstrual blood stem cell banking.