Key Instrumentation and techniques in Stem Cell Research (illustrated in Figure 2)
Microscopy Techniques
Microscopy is a cornerstone within stem cell biology as it enables the visualization of cellular structures, the monitoring of dynamic processes, and the analysis of molecular interactions. Fluorescence microscopy is a widely adopted method of choice because of its ability to visualize biomolecules and cells by tagging them with a fluorescent dye or fluorescent protein to identify a specific protein of interest, organelle, and other cellular components. This method is useful for the examination of morphology, cell fate specification (differentiation) and interactions with the stem cell niche (15). Confocal microscopy adds to fluorescence microscopy through the use of a pinhole aperture that eliminates out-of-focus light, producing sharper and more detailed images. This method is highly advantageous in producing the greatly sought-after high-resolution, three-dimensional reconstructions that stem cell researchers expect in studies of stem cells within their niches (16). Live-cell imaging allows researchers to observe stem cells as they behave in real-time, and visualize the STEM cell division, migration and differentiation under various spatially- and temporally-controlled conditions.
This step forward has transformed the field by enabling boxed visualization of subcellular structures and molecular complexes in stem cells and has led the way to better understanding the operating mechanisms in stem cell function and fate (17).
Flow Cytometry
To examine the physical and chemical properties of cell this is an analytical approach. With respect to stem cells, this technique serves as a highly effective tool for characterizing cellular populations based on surface and intracellular marker expression. Flow cytometry can quickly and quantitatively apply size, granularity, and fluorescence intensity to understand various cellular characteristics (18). Fluorescence-Activated Cell Sorting (FACS) can be used to separate required cell populations from the combination of multiple population based on fluorescence labelling. FACS is very useful for purifying stem cells and differentiating cells for further experiments, while allowing detailed investigation of defined cell types or developmental stages (19).
Mass Spectrometry
Mass spectrometry (MS) has become a significant tool in stem cell research for both proteomics and metabolomics. With MS, we can identify and quantify proteins, metabolites, and other biomolecules, and provide global profiles of stem cell states and states of response to varied stimuli. In proteomics, MS exposes differential protein composition as well as post-translational modifications and interactions amongst stem cells, and thus informs on cellular processes at the molecular level (20). Metabolomics uses MS to analyze the metabolic profiles of stem cells, providing insights into their metabolic states and pathways during differentiation and self-renewal processes (21).
Next-Generation Sequencing (NGS)
This technology has fundamentally transformed genomic and transcriptomic analysis in stem cell research.. NGS enables rapid and high-throughput sequencing of RNA and DNA, and has heightened resolution for examining the genetic and transcriptomic scopes pertinent to stem cell research. This technique allows for the identification of gene expression patterns, regulatory elements and genetic mutations that drive stem cell differentiation and behaviour (22). In addition, researchers can study the heterogeneity in stem cell populations by using NGS to carry out single-cell RNA sequencing, allowing the characterization of subpopulations that have unique functional attributes and molecular signatures (23).
High-Content Screening
It integrates automated imaging with sophisticated data analysis to enable large-scale screening of stem cells. HCS is used to analyze large populations of cells across differentiated conditions to obtain quantitative data on cellular phenotypes, morphology, and marker expression patterns. This technology is primarily applicable in drug development to identify compounds that improve or inhibit patterns of stem cell proliferation, differentiation, or survival (24). Due to its automated and scalable nature of HCS, this approach has become a very important tool for many studies, which include new discovery of therapeutic target and optimization of stem cell-based therapeutics.

Figure 2 : Key Instrumentation in Stem Cell Research

Stem Cell Culture & Maintenance
Culture Conditions and Media
The successful culture of stem cells relies on careful optimization of the culture conditions and media as shown in Table 3. The basic components of stem cell culture media are complementation of basal media with nutrients such as vitamins, amino acids, sodium, glucose, and growth factors and cytokines that promote cell proliferation and pluripotency. One of the most important supplements is serum, preferably fetal bovine serum (FBS), with a complex mix of growth factors, hormones, and adhesion factors. However, serum use can introduce variability and potential contamination risk, resulting in serum-free and chemically defined media (26). Chemically defined media create a controlled environment, which reduces variability and increases reproducibility in stem cell culture (27). Feeder layers usually consist of mitotically inactive murine embryonic fibroblasts (MEFs) which helps in the sustained proliferation of human embryonic stem cell populations (hESCs). Feeder layers also offer a supportive extracellular matrix and essential growth factor secretion, but the use of feeder layers from animals gave rise to potential cross species animal contamination and immunogenicity and resulted in the development of systems that do not need a feeder layer. Feeder-free culture systems use substrates such as Matrigel, which is a reconstituted basement membrane matrix, or synthetic matrices such as vitronectin and laminin and defined media to maintain pluripotency and promote stem-cell self-renewal (29). The developments have enabled the formation of xeno-free culture conditions that are fundamental for the translational clinical applications of therapies from stem-cell derivatives (27).
Cryopreservation and Thawing
Cryopreservation is a crucial approach for the long-term storage of stem cells, enabling researchers to preserve valuable cell lines and retain genetic and phenotypic stability over time (depicted in Table 3). Cryopreservation refers to the procedure of cooling the cells to sub-zero degrees, usually in liquid nitrogen (-196°C), while also introducing cryoprotective agents (CPAs) such as dimethyl sulfoxide (DMSO), and glycerol, which helps to protect cellular structures from the formation of ice crystals and prevent some degree of cellular damage during the freezing and thawing process (28). The most effective cryopreservation protocol utilizes a controlled-rate freezing procedure, which cools the cells slowly to help alleviate thermal shock and ice crystal formation (30). Thawing is also equally important and must also be performed quickly, to reduce the amount of time the cells are at intermediate temperatures and avoid the toxicity of the CPAs. Following thawing the cryovials are quickly placed into a 37°C water bath until thawed and subsequently, the CPAs are diluted to minimize cytotoxicity (31). The recovery and viability of post-thaw cells is greatly influenced by the thawing technique. Therefore, protocols must be meticulously optimized to promote viability and recovery, as well as for maintenance of cell functionality.
Passaging and Expansion
Passaging (also known as subculturing) is the move of cells from a dense culture vessel to a fresh culture vessel to allow more space for continued growth (see Table 3). Performing this procedure is critical to ensure that stem cells can remain viable and pluripotent for extended periods. Cells can be detached from the culture surface by enzymatic dissociation methods that utilize the enzymes trypsin or collagenase. However, these enzymes may damage cell surface proteins (marker proteins) which are critical to stem cell maintenance, which is why less damaging non-enzymatic dissociation solutions are used (32). Mechanical dissociation methods (e.g., scraping or pipetting) are an alternative but are generally less efficient and may induce cell stress than enzymatic methods. A "good" passaging protocol provides a balance between effectiveness of detaching cells from culture and minimizing affect on cell health. If more stem cells are to be expanded, particularly, if stem cells are to be produced at scale for potential therapeutic applications, suspension culture and bioreactor systems will be required. These systems must offer a controlled environment for expanding stem cells, while maintaining their undifferentiated state and functionality (33). Use of bioreactors has numerous advantages, namely with the ability to tightly control the culture environment, reduce human manipulation, and mechanise scaling up stem cell manufacture. Modulators of signalling pathways further support the maintenance of cell viability and pluripotency during passaging and expansion. ROCK inhibitors are one example of signalling pathway modulators which will enhance cell survival during single-cell dissociation of human pluripotent stem cells (hPSCs) since these cells are very sensitive to apoptosis when cultured as single cells (32). Advancements in techniques and technologies in cell culture is critical for reliable and reproducible expansion of stem cells so that they can be used for research and clinical application.

Differentiation Techniques
Protocols for Directed Differentiation
Directed differentiation procedures are important for differentiating pluripotent stem cells into discrete cell lineages that serve a prominent role in both basic research and in therapeutic applications. Directed differentiation procedures often recapitulate many of the natural developmental cues that occur during early embryonic development. In most approaches, researchers use a combination of signaling cues, bioactive compounds, and/or extracellular matrix components to direct stem cells through a series of developmental events. This sequence design is representative of both mesodermal induction and cardiac specification, similar to the process that occurs during these events in vivo. The application of regulatory growth cues and signal transduction pathways is an important component of the controlled differentiation strategy. Various regulatory growth mediators such as “fibroblast growth factor” (FGF), “epidermal growth factor” (EGF), and “transforming growth factor-beta” (TGF-β) are the primary categories of signalling molecules employed to promote stem cell differentiation into distinct cell types. As an example, differentiating neural progenitors from hPSCs requires the addition of FGF and EGF to regulate the proliferation and survival of neural precursors (34). Cell fate determination is influenced by various signaling pathways including Wnt, Notch, and Hedgehog, as precise regulation of these pathways can enhance differentiation efficiency and fidelity of differentiation protocols (35).
Organoid and 3D Culture Systems
The creation of organoid-based and three-dimensional culture platforms presents a major breakthrough in stem cell research that gives rise to more physiologically relevant models than conventional two-dimensional cultures. Organoids are three-dimensional, self-organizing constructions that are produced from stem cells and capture essential features of their respective organs. They provide an invaluable platform for investigating organogenesis, disease modelling, and drug screening with close resemblance to in vivo conditions (36). For instance, human embryonic stem cell-derived cerebral organoids were employed to simulate brain development and neurodevelopmental diseases and to elucidate the pathogenesis of diseases (36). These systems employ scaffolds composed of biocompatible materials like hydrogels to enable the growth and organization of stem cells into tissues with a high degree of complexity. Hydrogels may be engineered to mimic the biochemical and biomechanical characteristics of the extracellular matrix, to offer a conductive tissue environment for cell proliferation, differentiation, and the formation of tissue (37). Moreover, 3D bioprinting methods have been developed, which allow for the selective spatial organization of varied cell types and components of the extracellular matrix to create tissue-like tissue structures of high fidelity (38).
Co-culture Systems
Co-culture systems, where stem cells are maintained in combination with complementary cell populations, play a crucial role in regulating differentiation pathways and fostering the functional maturation of cells derived (as inferred in Table 3). Such systems mimic cell-cell interactions in vivo, delivering important signals impacting stem cell behavior. For example, co-culture of hPSCs with endothelial cells has been found to increase differentiation of hPSCs into functional hepatocytes, demonstrating the significance of cell-cell contact in liver organogenesis (39). Stromal feeder layers have been another form of frequent co-culture strategy. In hematopoietic stem cell (HSC) studies, co-culture of bone marrow-derived stromal cells is important to maintain HSCs in an undifferentiated state while promoting their cell fate to differentiate into blood cell lineages (40). Moreover, incorporation of microfluidic devices in co-culture systems makes it possible to have controlled cell-cell and cell-matrix interactions, which can be studied under dynamic conditions (41).

Genetic Engineering of Stem Cells
Editing of CRISPR/Cas9 and Genome
This technology has serving as a highly accurate, efficient, and adaptable platform for genome editing in stem cell biology. It consists of a guide RNA (gRNA), it guides the Cas9 endonuclease toward a DNA sequence to induce double-strand DNA breaks. These gaps are then repaired by the endogenous repair mechanisms of the cell, either through non-homologous end joining (NHEJ) or via homology-directed repair (HDR), allowing precise gene knockout, integration, or sequence modification (46). This technology has great importance in elucidating the function and regulation of genes in stem cells, enabling the creation of genetically matched cell lines for disease modeling and the development of novel therapeutic strategies. In stem cell science studies, CRISPR/Cas9 has been used to create disease models with the delivery of precise genetic mutations into human pluripotent stem cells (hPSCs). These models are priceless in the study of the pathophysiology of numerous genetic diseases and in screening for potential therapeutic agents (45). Furthermore, CRISPR/Cas9-based genome editing approaches applied to induced pluripotent stem cells (iPSCs) show strong promise in the context of precision medicine. For example, genetic disorders in iPSCs of patients suffering from monogenic diseases like cystic fibrosis or Duchenne muscular dystrophy have been corrected with success using this method (47).
Transfection and Transduction Methods
Effective delivery of genetic material into stem cells is crucial for genome editing and gene function analysis. Transfection and transduction processes include the delivery of DNA, RNA, or protein into cells. Viral delivery vehicles like lentiviruses and adenoviruses are also employed due to their efficient delivery capacity and stable genomic integration. Lentiviral vectors, specifically, are capable of efficiently delivering genetic material into both quiescent and proliferating cells, and are thus amenable to a wide variety of stem cell types (48). Non-viral delivery systems also include a range of alternative approaches such as electroporation, lipofection, and nanoparticle-mediated delivery, which offer distinct advantages as substitute strategies. Electroporation comprises the use of an electric field for transient permeabilization of the cell membrane, thereby facilitating the entry of nucleic acids into the cell. This process is very efficient but is cytotoxic in nature, thus requiring optimization of parameters to get the balance between efficiency and cell viability (49). Lipofection utilizes a lipid-mediated delivery system that complexes with nucleic acids, allowing the nucleic acids to be taken up by cells by endocytosis. Although lipofection is less efficient compared to viral approaches, lipofection is not immunogenic and carries no danger of insertional mutagenesis (50). Nanoparticle-delivery, by materials like gold nanoparticles or polymeric nanoparticles, is an emerging area that provides exact control of delivery and release kinetics (51).
Induced Pluripotent Stem Cells (iPSCs)
It represents a landmark advancement within regenerative medicine, enabling the directed conversion of mature somatic cells to pluripotency. It entails the induced activation of key transcriptional regulators—commonly OCT4, SOX2, KLF4, and c-MYC—into adult cells, which are then reprogrammed into a state resembling embryonic stem cells (52). iPSCs are also able to give rise to all differentiated cell lineages, which positions them as a substantial reservoir of patient-derived cells for therapeutic and research purposes. Reprogramming somatic cells into a pluripotent state has profound implications in disease modeling, therapeutic discovery, and precision medicine.. Patient-specific iPSCs can be directed to differentiate into pathology-relevant cell types, and thereby underlying disease processes can be studied in the patient-specific background. For instance, iPSCs derived from individuals affected by neurodegenerative disorders, such as Parkinson’s disease, can be induced into dopaminergic neurons to characterize disease pathology and screen potential therapeutic interventions (45) Moreover, the employment of iPSCs circumvents ethical concerns associated with embryonic stem cell use and reduces the risk of immunological rejection in cellular transplantation therapies. Reprogramming and maintenance of iPSCs demand strict culture conditions in order to preserve their pluripotency and genomic integrity. Improvements in reprogramming methods, such as the application of non-integrating vectors and small molecules, have enhanced the safety and efficacy of iPSC generation (53). Such progress is essential in taking iPSC technology from the laboratory bench to the bedside, where safety and consistency are the priority.
Advanced Imaging and Analysis Techniques
Single-Cell Analysis
This method is now an essential tool for stem cell research since it dissects heterogeneity of the cell and detects rare cell populations that may be "hidden" in bulk analyses, as outlined in Table 5. FACS allows the separation of single cells on the basis of expression of a particular marker, offering the isolation into separate subpopulations for examination (54). Microfluidic devices utilize the manipulation of fluids in small volumes with high precision and efficiency for the capture of individual cells from complicated tissues. Such platforms are able to combine several steps, such as cell isolation, lysis, and amplification of nucleic acids, into a single chip, and thus are better for high-throughput single-cell analysis (55). Single-cell RNA sequencing transformed the comprehension of cell diversity and dynamics of stem cell populations. Through the profiling of transcriptomes in single cells, scRNA-seq gives knowledge about gene expression heterogeneity and additional recognition of cell states and lineages. This method has been utilized to profile the development paths in embryonic stem cells, revealing transitional states and lineage bifurcations which are not obvious under bulk RNA sequencing (56). Another step forward in single-cell multi-omics is the combined application of transcriptomic data with proteomic or epigenomic profiling at single-cell resolution, providing comprehensive views of cellular states and regulatory architectures, further facilitating understanding of stem cell biology(57).
Live-Cell Tracking and Imaging Technique
Live-cell imaging and tracking methods are important to investigate the dynamics of multiple cellular events in stem cells, such as migration, division, as well as lineage differentiation, in real time (Table 5). Time-lapse microscopy is a popular technique and consists of taking a sequence of images at set intervals so that researchers can track changes in cell behaviour and morphology with the passage of time. This method can be merged with fluorescent probes to enable the visualization of certain proteins or organelles and yield information regarding intracellular processes and signalling events (58). Sophisticated live-cell imaging methods, including Förster resonance energy transfer (FRET) and fluorescence lifetime imaging microscopy (FLIM), allow detailed examination of molecular interactions and signaling behavior in live cells. FRET quantifies energy transfer between two fluorophores that are near each other to uncover protein-protein interactions and conformational changes (59). While FLIM quantifies fluorescence decay rate, giving information on fluorescent molecule local environment, e.g., pH or ion concentration (60). These methods are most useful for investigating spatiotemporal signalling pathway dynamics in stem cells, and they help us understand the regulation of extracellular signals and intracellular networks controlling stem cell fate choices.
Biophysical Techniques
This technique approaches provide the potential capabilities to probe the mechanical properties and stem cell interactions at a nanoscale (Table 5). AFM uses a sharp-tipped cantilever to scan the cell surface, allowing high-resolution topographical images, and determinations of mechanical properties of stiffness and elasticity (61). This method has been applied to examine the biomechanical characteristics associated with stem cell niches and how mechanical cues regulate stem cell behavior and differentiation(62). Optical tweezers have very tightly focused laser beams with which to trap and move around microscopic objects, and are used to quantify forces and displacements accurately. This method is primarily beneficial for the investigation of interactions between cells and between cells and the extracellular matrix, and for the controlled manipulation of single molecules to assess their mechanical properties and interactions. (63). Magnetic tweezers, in which controlled magnetic forces are applied to magnetic beads coupled to molecules or cells, provide another means of investigating the mechanical interactions and mechanical transduction pathways in stem cells (64). These sophisticated biophysical methods are important in uncovering how the physical forces and mechanical properties affect stem cell function. It also gives a better insight into the mechanobiology of stem cells, emphasizing the interaction between the mechanical and biochemical signals in controlling stem cell behaviour and fate.

Biomaterials and Scaffolds
Synthetic and Natural Biomaterials
The biomaterials field has immensely contributed to stem cell research by providing different innovative materials for supporting cell culture, differentiation, and tissue engineering. Synthetic as well as natural biomaterials have very important functions in providing conditions that simulate the native extracellular matrix (ECM), necessary to maintain stem cell functionality and guide their differentiation. Synthetic biomaterials like polylactic acid (PLA), polyglycolic acid (PGA), and polycaprolactone (PCL) offer the benefit of mechanical properties, biodegradability, and the potential for precise engineering to tissue needs (65). They can be processed into fibers, meshes, and hydrogels to support structure and be altered in terms of surface modification for cell attachment and growth. Natural biomaterials, which are extracted from biological sources, are similar in composition and properties to the ECM and present a more physiologically similar environment for stem cells. Some examples include collagen, gelatin, alginate, and chitosan, all of which are extensively employed in stem cell research. Collagen, the largest protein present in the ECM, is involved in the structural integrity and aids in cell adhesion and proliferation. It has been employed to design scaffolds that support the directed differentiation of stem cells toward diverse lineages, including osteogenic and chondrogenic pathways (66). Alginate from brown seaweed is employed to encapsulate stem cells and offer a three-dimensional (3D) culture milieu that promotes intercellular interactions and enhances lineage differentiation (67).
The combination of natural and synthetic biomaterials will leverage the strengths of both to develop a hybrid scaffolds with mechanical strength and biological activity. For example, hybrid scaffolds made from PCL blended with gelatin or collagen have demonstrated enhanced biocompatibility and promotion of stem cell differentiation (67). These developments in biomaterials are critical for the establishment of effective stem cell therapies and tissue engineering products, to choose a particular biomaterial to impact outcomes dramatically.
3D Bioprinting
Three-dimensional bioprinting represents an advanced technological approach for tissue engineering that allows building complex tissue structures with exact precision by layer-by-layer depositing cells and biomaterials. The method has the capability to create highly ordered, functional tissues that will precisely simulate natural organs, giving a unique tool to regenerative medicine and disease modeling. 3D bioprinting has the versatility to use different bioinks made up of living cells dispersed in a biomaterial matrix. These bioinks may be optimized for particular biochemical and mechanical stimuli that promote cell viability, growth, and differentiation (68).
Three-dimensional bioprinting holds considerable promise for tissue engineering applications. Due to the capability of controlling the spatial organization of cells and biomaterials with high accuracy, 3D bioprinting has the ability to form tissue constructs with heterogeneous compositions that are as complex as native tissues. Bioprinted cardiac patches with cardiomyocytes, fibroblasts, and endothelial cells, for instance, have shown synchronized beating and vascularization and thus the potential to repair cardiac tissue (69). Along the same lines, bioprinted liver constructs are proving to be useful in liver function and disease research, providing an experimental framework for pharmacological screening and toxicity assessment (69). In addition, development of bioprinting technologies like multi-material printing and the integration of microfluidic systems is widening the scope of the technique. Multi-material bioprinting enables the co-deposition of various cell types and biomaterials simultaneously, creating more sophisticated and functional tissue constructs. The incorporation of microfluidic channels in bioprinted tissues has the ability to mimic blood vessels, which further facilitates nutrient and oxygen supply to cells and tissue viability and function (66). 3D bioprinting holds the promise to go beyond tissue engineering and extend to precision medicine. Using cells derived from individual patients, tissues bioprinted can be tailored to match the patient’s genetic and physiological profile, minimizing the likelihood of immune rejection and maximizing the therapeutic efficacy of regenerative therapy. This patient-tailored therapy can transform the therapy of numerous conditions, ranging from organ failure to genetic disorders, by allowing tailored tissue replacements and therapy models (70).
Applications and Future Directions
Clinical Applications and Trials
Stem cell clinical applications are leading the regenerative medicine field, providing potential therapies for numerous conditions with limited therapeutic treatments available. Therapies based on stem cells seek to restore or substitute damaged tissues and organs by taking advantage of stem cell differentiation capabilities into different cell types. One of the most consolidated uses is the hematopoietic stem cell transplantation (HSCT) as a treatment for blood diseases like leukemia, lymphoma, and some genetic disorders (71). The therapy has been optimized over decades and is considered a paradigm for other stem cell therapies.
More recent developments utilize the mesenchymal stem cells (MSCs) for the treatment of inflammatory and autoimmune diseases. MSCs, with their immunomodulatory capacity, have been promising in clinical trials of diseases like Crohn's disease, graft-versus-host disease (GVHD), and multiple sclerosis (69). The ability of these cells to modulate immune responses and induce tissue repair makes them a potential candidate of therapeutic agents in a range of inflammatory situations. The derivation of induced pluripotent stem cells (iPSCs) provided a novel advancement for personalized medicine. iPSCs, which are generated from an individual’s own cells, can be directed to differentiate into any cell lineage, and will reduce the likelihood of immune rejection. Clinical trials are investigating the use of retinal cells generated from iPSCs to treat age-related macular degeneration (AMD) as well as dopaminergic neurons to treat Parkinson's disease (71). These experiments are the zenith of regenerative medicine that seek to offer long-term cures for degenerative diseases.
Stem Cells in Disease Modelling
Stem cells are also vital in disease modelling, as they provide additional information about the pathophysiology of diseases and allow us to develop new therapies. Human pluripotent stem cells (hPSCs), such as embryonic stem cells (ESCs) and iPSCs, can be differentiated into cell types relevant to disease, to generate an in vitro model that reflects the disease condition. These models are especially useful for the investigation of genetic diseases because patient-specific iPSCs are able to produce cell types undergoing the disease, enabling detailed examination of disease-related mechanisms and the evaluation of candidate therapeutic compounds (72). One of the key uses of stem cell–based disease modeling lies in neurodegenerative disorders, including Alzheimer’s disease, Parkinson’s disease, and amyotrophic lateral sclerosis (ALS). iPSC-derived neurons of these patients have already been employed to analyze disease-specific cellular phenotypes of protein aggregation, mitochondrial impairment, and synaptic deficits (73). These models allow the establishment of a high-throughput drug screening framework, in order to find compounds that can reverse disease phenotypes and possibly result in novel treatments. Aside from neurodegenerative conditions, stem cell models are also being applied to model cardiac diseases, diabetes, and different cancers. For instance, iPSC-derived cardiomyocytes are used to model genetic arrhythmias of the heart, offering future prospective into the underlying electrophysiological deficits and evaluating the effectiveness of anti-arrhythmic agents (74). Likewise, stem cell-derived pancreatic beta cells are also utilized to study diabetes as well as evaluate novel insulin-sensitizing drugs.
Ethical and Regulatory Considerations
The progress of stem cell science and its translation into the clinical setting presents with the importance of the implication of ethical and regulatory concerns. The ethical concerns mostly constitute the origin of stem cell sources, such as human, which involves its destruction. These have been the subjects of controversial discussions and inconsistent regulations across nations. Conversely, the generation of iPSCs has mitigated some of the ethical issues, as they are no longer needed to utilize embryos and can be derived from somatic cells in adults (75). The regulatory systems are largely necessitated for ensuring the safety and therapeutic effectiveness of stem cell therapies. The regulatory agencies, establish core guidelines governing the clinical translation of stem cell therapies. The standards include strict preclinical testing requirements, good manufacturing practices, and clinical trial design for the assurance of safety and efficacy of stem cell products in humans (76). Nonetheless, the quick momentum in stem cell innovation is ahead of the regulation capacity, which requires continuous updates and revisions to counter new challenges and innovations. Besides the regulatory aspects, ethical concerns related to patients' consent, privacy, and genetic information utilization need to be properly managed. Informed consent procedures ensure patients are well-informed of the possible benefits and risks of being part of a stem cell-based clinical study or clinical trials. Furthermore, utilization of genetic data derived from stem cell research also raises issues of privacy of data and the risk of genetic discrimination, thus necessitating strong data protection protocols (77).