Cell Transplantation Techniques Research Articles

Mechanical stretch and chronotherapeutic techniques for progenitor cell transplantation and biomaterials

Mechanical stretch and chronotherapeutic techniques for progenitor cell transplantation and biomaterials

Identification and Manipulation of Spermatogonial Stem Cells with the Aim of Inducing Spermatogenesis in Vitro.

Assisted reproduction techniques for infertile men with non-obstructive azoospermia require a sufficient number of functional germ cells produced in vitro. Understanding the mechanisms that allow the resumption of spermatogenesis outside the testicular environment is crucial for fertility preservation in these patients. A review of the literature was conducted using databases such as PubMed, Scopus and Web of Science, with keywords including "spermatogonial stem cell," "germ cells," "male factor infertility," and "enrichment and propagation of SSCs in vitro." Currently, two models-"in vivo" and "in vitro"-have been developed for producing haploid germ cells. The "in vivo" models include spermatogonial stem cell transplantation and testicular xenograft techniques. In contrast, the "in vitro" models consist of conventional culture systems, organ culture, and three-dimensional culture systems, all designed to induce spermatogenesis in vitro. These culture systems enable the simulation of the seminiferous epithelium in vitro, which facilitates better regulation of cell-signaling pathways that control the self-renewal and differentiation of SSCs. This review provides up-to-date information on the organization of SSCs, focusing on the identification, proliferation, and differentiation of spermatogonia in vitro.

Research progress of stem cell therapy for neurological diseases

Neurological diseases are considered one of the leading causes of death and disability worldwide; not only do patients suffer, but they also place a significant economic burden on families and society. Traditional medications now show limited efficacy, and surgical interventions can be dangerous. The need to develop new medicines is critical as more people become aware of the immense burden this disease imposes on society and the limited number of viable therapeutic options that currently exist. Modern research is increasingly focusing on novel and potent approaches to treat neurological illnesses to address these challenges. One promising method is the stem cell-based therapy. The development of stem cell transplantation techniques and cellular therapies in recent years has provided new hope for the treatment of neurological disorders. Stem cell therapies have demonstrated therapeutic potential in animal models, and different types of stem cells have been used in clinical trials. In this study, we explored the applications of mesenchymal stem cells, dental pulp stem cells, and induced pluripotent stem cells in neurological diseases. In recent years, we have examined the current status and progress of stem cell applications in treating neurological diseases, both domestically and internationally, and reviewed some of the key challenges encountered.

Utilizing Multifaceted Approaches to Target Drug Delivery in the Brain: From Nanoparticles to Biological Therapies.

The blood-brain barrier (BBB) poses an important obstacle to treating neurological disorders because it limits the entry of therapeutic agents into the central nervous system (CNS). Surmounting this barrier is crucial for delivering drugs effectively and targeting precise areas of the brain affected by conditions like Parkinson's disease, Alzheimer's disease, and brain tumors. This review examines the diverse strategies employed to enhance brain targeting, including nanotechnology, viral vectors, and biological therapies. Nanoparticles, liposomes, and dendrimers offer promising approaches for encapsulating drugs and facilitating their transport across the BBB. Viral vectors, such as adeno-associated viruses, demonstrate high transfection efficiency for gene therapy applications in CNS diseases. Biological therapies, including stem cell transplantation and neuromodulation techniques, can potentially restore normal cellular function and treat genetic disorders. Challenges such as BBB permeability, safety concerns, and regulatory considerations are discussed, along with future perspectives on precision medicine, noninvasive delivery methods, and biomarker discovery. By addressing these challenges and embracing innovative approaches, the field of brain drug targeting aims to transfer the way that neurological illness is treated and improve patient outcomes.

Femtosecond Laser-Assisted Autologous Glueless Simple Limbal Epithelial Transplantation in Unilateral Limbal Stem Cell Deficiency: 12-Month Outcome of the First Clinical Cases.

Surgical treatment of unilateral limbal stem cell deficiency (LSCD) is based on limbal stem cell transplantation. Glueless simple limbal epithelial transplantation (G-SLET) technique implements several limbal micrografts harvested from the healthy eye of the same patient into the peripheral corneal tunnels without the use of fibrin glue and human amniotic membrane. A novel customized algorithm and software for a low-energy femtosecond laser (FSL) were developed and tested using 5 pairs of isolated porcine eyes. FSL-assisted G-SLET modification was assessed in 3 clinical cases of unilateral LSCD caused by chemical burns. Corneal epithelization efficacy, best-corrected visual acuity, corneal epithelial mapping, central corneal thickness, and impression cytology with immunohistochemical examination were evaluated. All patients were followed up for 12 months postoperatively. The FSL set for 100% energy and the pattern of 8 nonpenetrating vertical cuts with oblique tunnel portions having variable incision depths and diameter of 8.5 mm and higher were selected for further clinical evaluation. Clinically, stable corneal epithelialization was achieved 2 to 3 weeks after intervention. At the 6-month follow-up, all patients had a healthy corneal epithelium with limbal micrografts visible inside the corneal tunnels. Best-corrected visual acuity markedly improved in 2 cases, but not in the third case with severe corneal stromal scarring. In addition, all patients noted a full-scale reduction in subjective complaints and substantial improvement in their quality of life. The FSL-assisted G-SLET is a new technique for autologous limbal stem cell transplantation in patients with unilateral LSCD. It allows the standardization of corneal tunnel localization and dimensions, thereby increasing the safety of the surgical procedure.

Exploring the Potential of Skin Cell Culture and the Vital Role of Skin Grafts in Regenerative Medicine and Biology

The skin is acknowledged as the biggest organ, covering the exterior of the body and serving as a physical barrier. The integumentary system is made up of three layers of tissue: the epidermis, dermis, and hypodermis. For transplantation, epidermal cell cultures have been around for a while. The epidermal stem cells can proliferate and self-renew in a manner that allows for rapid healing since they are unipotent, meaning they can only produce one type of cell. In therapeutic terms, cultured keratinocytes have been described as an autologous cell-based therapy that provides a source of autologous tissue, reducing the need for large donor sites and minimizing the risk of immune rejection. This review explores the composition and functioning of the skin’s extracellular matrix, along with strategies to incorporate ECM components into the cell culture techniques. Rebuilding skin flaws without consideration for side effects is skin grafting. The tissue lacks its circulatory supply and must receive it from the bed in which it is inserted. The skin cells have contributed to developing skin graft and tissue engineering approaches for repairing damaged skin. Similar to grafting, the skin graft is divided into sections based on the quantity of dermis involved: split-thickness skin and full-thickness skin. Transplanting autologous keratinocytes is an alternative to autologous skin grafting. In the wound bed, epidermal stem cells encourage angiogenesis. After invading the fibrin wound clot, angiogenic capillary sprouts form a microvascular network throughout the granulation tissue in a few days. In this article, authors will review the current methods and application of skin cell culture for transplantation including different cell sources, culture conditions, and transplantation techniques. The future direction of this approach is in the context of regenerative medicine and tissue engineering.

Research progress of dental pulp regeneration treatment.

The dental pulp is the only soft tissue structure within the tooth, serving functions such as sensation and nutrition. However, the dental pulp is highly susceptible to necrosis due to external factors. Currently, root canal therapy is the most commonly used treatment for pulp necrosis. Nevertheless, teeth treated with root canal therapy are prone to secondary infections and adverse outcomes like vertical root fractures. Regenerative endodontic therapy has emerged as a solution, aiming to replace damaged tooth structures, including dentin, root structure, and the pulp-dentin complex cells. This approach demonstrates significant advantages in addressing clinical symptoms and achieving regeneration of the root and even the pulp. Since the discovery of dental pulp stem cells, regenerative endodontic therapy has gained new momentum. Advances in cell transplantation and cell homing techniques have rapidly developed, showing promising potential for clinical applications.

A Challenging Case of Simple Limbal Epithelial Transplantation (SLET) in Limbal Stem Cell Deficiency Following Chemical Injury: A Case Report

Introduction : Limbal stem cell deficiency (LSCD) often arises following trauma or inflammation that hinders corneal epithelium healing, commonly caused by chemical or thermal injuries. Despite significant advancements, treating LSCD remains challenging. This case highlights a challenging case of simple limbal epithelial transplantation (SLET) in LSCD following chemical injury.
 Case Illustration : A 20-year-old male presented with reduced vision in left eye following an alkaline chemical injury (grade III) was diagnosed with unilateral LSCD. Upon examination, conjunctivalization of the peripheral cornea, a central corneal cicatrix, and stromal neovascularization present after previous necrotomy, tenoplasty, and amniotic membrane transplantation (AMT). The patient then underwent SLET, with limbal biopsies from the collateral eye. Four months after surgery the visual acuity improved from 3/60 to 6/15 with spectacles. However, the cornea conjunctivalization still developed despite improved visual acuity.
 Discussion : To effectively manage the recurrence of stem cell deficiency, other stem cell transplantation techniques are needed to manage the problem. Combining different limbal stem cell transplantation approaches can be customised to address the problem and achieve lasting results.
 Conclusion : Prompt diagnosis and treatment of LSCD is crucial to prevent complications and blindness. SLET is a surgical option for LSCD after chemical injury, but this report highlights a challenging case and the need for the alternatives approaches.

Fish germ cell cryobanking and transplanting for conservation.

The unprecedented loss of global biodiversity is linked to multiple anthropogenic stressors. New conservation technologies are urgently needed to mitigate this loss. The rights, knowledge and perspectives of Indigenous peoples in biodiversity conservation-including the development and application of new technologies-are increasingly recognised. Advances in germplasm cryopreservation and germ cell transplantation (termed 'broodstock surrogacy') techniques offer exciting tools to preserve biodiversity, but their application has been underappreciated. Here, we use teleost fishes as an exemplar group to outline (1) the power of these techniques to preserve genome-wide genetic diversity, (2) the need to apply a conservation genomic lens when selecting individuals for germplasm cryobanking and broodstock surrogacy and (3) the value of considering the cultural significance of these genomic resources. We conclude by discussing the opportunities and challenges of these techniques for conserving biodiversity in threatened teleost fish and beyond.

BONE MARROW-DERIVED STEM CELL TRANSPLANTATION FOR OSTEOCHONDRAL DEFECTS OF THE KNEE: A PROSPECTIVE CLINICAL STUDY

AbstractBackgroundAutologous chondrocyte implantation is a NICE approved intervention however it involves the morbidity of two operations, a prolonged rehabilitation and substantial healthcare costs. This study describes a novel, one-step, bone marrow (BM) derived mesenchymal stem cell (MSC) transplantation technique for treating knee osteochondral lesions and presents our prospective clinical study investigating the success of this technique in 206 lesions over a 5 year period.MethodologyThe surgical technique involves harvesting BM from patients’ anterior superior iliac spines, centrifugation to isolate MSCs and seeding into a type 1 collagen scaffold (SyngenitTM Biomatrix). Autologous fibrin glue is used to secure the scaffold into the defect. Inclusion criteria included patients aged 15 – 55 years old with symptomatic osteochondral lesions >1cm2. Exclusion criteria included patients with ligament instability, uncorrected alignment, inflammatory arthropathy and a Body Mass Index >35 kg/m2. Outcome measures included the Modified Cincinnati Knee Rating System (MCKRS), complications and reoperations.ResultsMean MCKR scores showed statistically significant improvements compared to pre-operative scores at 6 months 58.79 ± 3.5 and 1 year postoperatively 63.82 ± 3.93 with further improvements at 2 years and 5 years which did not reach statistical significance. Survival rates were 97.9%, 94% and 93.2% at 1, 2 and 5 years. Multiple regression analysis identified previous cartilage surgery, microfracture and age as factors affecting MCKRS scores (p < 0.029, 0.001 and 0.030, respectively).ConclusionsOne-step BM derived stem cell transplantation demonstrates satisfactory outcomes over a 5 year period.

Tooth-derived stem cells integrated biomaterials for bone and dental tissue engineering.

Recent years have seen the emergence of tissue engineering strategies as a means to overcome some of the limits of conventional medical treatment. A biomaterial with tailored physio-chemical characteristics is used in this sophisticated method to transport stem cells and growth factors/bioactive substances, or to attract local endogenous cells, enabling new tissue formation. Biomaterials might serve as a biomimetic structure inspired by the natural milieu, assisting the cells in establishing their natural relationships. Such a method would benefit from having ready access to an abundant reservoir of stem cells with strong tissue regeneration capacity, in addition to using biological compatible material to promote new tissue creation. Teeth may have a plethora of self-renewing, multipotent mesenchymal stem cell (MSC) populations. Recent advancements and promising directions for cell transplantation and homing techniques using dental MSCs for tissue regeneration are discussed in this review paper. Overall, this research paints a picture of the present landscape of new approaches to using tooth-derived MSCs in conjunction with biomaterials and bioactive substances for tissue regeneration.

Pulp-Dentin Complex Regeneration with Cell Transplantation Technique Using Stem Cells Derived from Human Deciduous Teeth: Histological and Immunohistochemical Study in Immunosuppressed Rats.

The aim of this study was to histologically verify the performance of pulp-derived stem cells used in the pulp-dentin complex regeneration. Maxillary molars of 12 immunosuppressed rats were divided into two groups: the SC (stem cells) group, and the PBS (just standard phosphate-buffered saline) group. After pulpectomy and canal preparation, the teeth received the designated materials, and the cavities were sealed. After 12 weeks, the animals were euthanized, and the specimens underwent histological processing and qualitative evaluation of intracanal connective tissue, odontoblast-like cells, intracanal mineralized tissue, and periapical inflammatory infiltrate. Immunohistochemical evaluation was performed to detect dentin matrix protein 1 (DMP1). In the PBS group, an amorphous substance and remnants of mineralized tissue were observed throughout the canal, and abundant inflammatory cells were observed in the periapical region. In the SC group, an amorphous substance and remnants of mineralized tissue were observed throughout the canal; odontoblasts-like cells immunopositive for DMP1 and mineral plug were observed in the apical region of the canal; and a mild inflammatory infiltrate, intense vascularization, and neoformation of organized connective tissue were observed in the periapical region. In conclusion, the transplantation of human pulp stem cells promoted partial pulp tissue neoformation in adult rat molars.

Development of stem cell therapy for atherosclerosis.

Cardiovascular disease (CVD) has a high incidence and low cure rate worldwide, and atherosclerosis (AS) is the main factor inducing cardiovascular disease, of which lipid deposition in the vessel wall is the main marker of AS. Currently, although statins can be used to lower lipids and low-density lipoprotein (LDL) in AS, the cure rate for AS remains low. Therefore, there is an urgent need to develop new therapeutic approaches, and stem cells are now widely studied, while stem cells are a class of cell types that always maintain the ability to differentiate and can differentiate to form other cells and tissues, and stem cell transplantation techniques have shown efficacy in the treatment of other diseases. With the establishment of cellular therapies and continued research in stem cell technology, stem cells are also being used to address the problem of AS. In this paper, we focus on recent research advances in stem cell therapy for AS and briefly summarize the relevant factors that induce the formation of AS. We mainly discuss the efficacy and application prospects of mesenchymal stem cells (MSCs) for the treatment of AS, in addition to the partial role and potential of exosomes in the treatment of AS. Further, provide new ideas for the clinical application of stem cells.

Cost-Effectiveness of Unrelated Umbilical Cord Blood Transplantation versus HLA-Haploidentical Related Bone Marrow Transplantation: Evidence from BMT CTN 1101

BMT CTN 1101 was a Phase III randomized controlled trial comparing reduced-intensity conditioning followed by double unrelated umbilical cord blood transplantation (UCBT) versus HLA-haploidentical related donor bone marrow transplantation (haplo-BMT) for patients with high-risk hematologic malignancies. Here we report the results of a parallel cost-effectiveness analysis of these 2 hematopoietic stem cell transplantation (HCT) techniques. In this study, 368 patients were randomized to unrelated UCBT (n=186) or haplo-BMT (n=182). We estimated healthcare utilization and costs using propensity score-matched haplo-BMT recipients from the OptumLabs Data Warehouse for trial participants age <65 years and Medicare claims for participants age ≥65 years. Weibull models were used to estimate 20-year survival. EQ-5D surveys by trial participants were used to estimate quality-adjusted life-years (QALYs). At a 5-year follow-up, survival was 42% for haplo-BMT recipients versus 36% for UCBT recipients (P=.06). Over a 20-year time horizon, haplo-BMT is expected to be more effective (+.63 QALY) and more costly (+$118,953) for persons age <65 years. For those age ≥65 years, haplo-BMT is expected to be more effective and less costly. In one-way uncertainty analyses, for persons age <65, the cost per QALY result was most sensitive to life-years and health state utilities, whereas for those age ≥65, life- years were more influential than costs and health state utilities. Compared to UCBT, haplo-BMT was moderately more cost-effective for patients age <65 years and less costly and more effective for persons age ≥65 years. Haplo-BMT is a fair value choice for commercially insured patients with high-risk leukemia and lymphoma who require HCT. For Medicare enrollees, haplo-BMT is a preferred choice when considering costs and outcomes.

Cell transplantation techniques for cartilage restoration

BackgroundFocal articular chondral lesions are a common finding among patients presenting with knee pain. If symptomatic and unresponsive to conservative treatment, cell transplantation techniques offer a unique solution to address larger defects by engineering chondrocytes to integrate within the subchondral bone of the lesion and regenerate cartilage. ObjectivesThe purpose of this article is to review the evaluation of, and clinical decision making for, patients being considered for cell transplantation and the available techniques an orthopaedic surgeon has at their disposal. MethodsA review of recent literature regarding cartilage defects in the knee and cell transplantation techniques was performed to provide strategies for evaluating and treating chondral defects with matrix-induced autologous chondrocyte implantation (MACI) or particulated juvenile allograft cartilage (PJAC) procedures. ResultsChondral defects in the knee can be treated with patients’ own chondrocytes embedded into a collagen membrane as a MACI procedure, or minced autologous donor cartilage that is then implanted onto a scaffold as a PJAC procedure. These cell transplantation techniques offer advantages compared to bone marrow stimulation or mosaicplasty, and have shown clinically significant improvements in outcome scores with low rates of complications. ConclusionCell transplantation techniques such as MACI and PJAC offer treatment options that can effectively address large full-thickness chondral defects in the tibiofemoral joint or patellofemoral joint that may respond poorly to bone marrow stimulation or mosaicplasty.

Fabrication of nanofibrous mat surrounded hydrogel scaffold as an encapsulation device for encapsulating pancreas β cells

The main barriers to cells or organ transplantation such as pancreatic β-cells are the need for lifelong immune suppression and the shortage of donors. It may be overcome via cell encapsulation and transplantation techniques. Hydrogels provide a suitable ECM-like microenvironment for cells to adhere, survive, and function, while weakly performing as an immune barrier. In this study, we aimed to macro-encapsulate islet cells in a dual encapsulation device with collagen hydrogel and PCL nanofiber to provide an immune-isolated environment for cells to function more efficiently, where immune cells are not allowed to enter but oxygen, insulin, and nutrients can pass through. PCL thin mats with the pores diameter of 500 nm were synthesized by electrospinning and characterized by scanning electron microscope, porosity measurement, tensile strength test, and contact angle measurement. Collagen hydrogel was fabricated by extracting collagen fibers from rat tail tendons and solving them in acetic acid. β-cells (CRI-D2 cell line) encapsulated after neutralizing collagen solution (pH ≈ 7.4). Cell-collagen gel complex was poured into the nanofibrous mat packets to fabricate the whole device. Histology evaluation, cell viability, and cell function tests were done in 10 days. Live/dead assay of Cri-D2 cells encapsulated within the device showed that cells have diffuse distribution at the core of the hydrogel and the device. Also, cluster formation was seen and shows these cells can live in groups. To identify cells’ function within the device in these 10 days samples’ supernatant insulin level was measured by chemiluminescent immunoassay. It just showed a positive result for existing insulin within the medium. Based on our results, this device presents adequate features to be a good immune-isolation device for cell transplanting.

A Review of Acute and Long-Term Neurological Complications Following Haematopoietic Stem Cell Transplant for Paediatric Acute Lymphoblastic Leukaemia.

Despite advances in haematopoietic stem cell transplant (HSCT) techniques, the risk of serious side effects and complications still exists. Neurological complications, both acute and long term, are common following HSCT and contribute to significant morbidity and mortality. The aetiology of neurotoxicity includes infections and a wide variety of non-infectious causes such as drug toxicities, metabolic abnormalities, irradiation, vascular and immunologic events and the leukaemia itself. The majority of the literature on this subject is focussed on adults. The impact of the combination of neurotoxic drugs given before and during HSCT, radiotherapy and neurological complications on the developing and vulnerable paediatric and adolescent brain remains unclear. Moreover, the age-related sensitivity of the nervous system to toxic insults is still being investigated. In this article, we review current evidence regarding neurotoxicity following HSCT for acute lymphoblastic leukaemia in childhood. We focus on acute and long-term impacts. Understanding the aetiology and long-term sequelae of neurological complications in children is particularly important in the current era of immunotherapy for acute lymphoblastic leukaemia (such as chimeric antigen receptor T cells and bi-specific T-cell engager antibodies), which have well-known and common neurological side effects and may represent a future treatment modality for at least a fraction of HSCT-recipients.

Corneal Epithelial Stem Cells-Physiology, Pathophysiology and Therapeutic Options.

In the human cornea, regeneration of the epithelium is regulated by the stem cell reservoir of the limbus, which is the marginal region of the cornea representing the anatomical and functional border between the corneal and conjunctival epithelium. In support of this concept, extensive limbal damage, e.g., by chemical or thermal injury, inflammation, or surgery, may induce limbal stem cell deficiency (LSCD) leading to vascularization and opacification of the cornea and eventually vision loss. These acquired forms of limbal stem cell deficiency may occur uni- or bilaterally, which is important for the choice of treatment. Moreover, a variety of inherited diseases, such as congenital aniridia or dyskeratosis congenita, are characterized by LSCD typically occurring bilaterally. Several techniques of autologous and allogenic stem cell transplantation have been established. The limbus can be restored by transplantation of whole limbal grafts, small limbal biopsies or by ex vivo-expanded limbal cells. In this review, the physiology of the corneal epithelium, the pathophysiology of LSCD, and the therapeutic options will be presented.

Magic-TT Mediated Hematopoietic Stem/Progenitor Cells Promote Hematopoietic Recovery after Autologous Bone Marrow Transplantation

Background: Hematopoietic stem cell transplantation (HSCT) was widely used. Rapid and effective reconstruction would directly affect the effects of transplantation (including survival rate, blood routine examination and implantation rate of donor cells). In previous research (Blood 2015 126:5404), we developed a novel cell transplantation technique named magnetism-induced cell target transplantation (MagIC-TT). The purpose of this research is to explore whether MagIC-TT could promote hematopoietic recovery after autologous bone marrow transplantation.Methods: 1) 66 female C57BL/6 mice with the condition of specific pathogen free (SPF) were randomly divided into MagIC-TT group and non-MagIC-TT group. All recipient mice were irradiated by 7.5 Gy X-ray one day before transplantation. 2) eGFP+ bone marrow hematopoietic stem/progenitor cells (HS/PCs) were flushed out from the femur, tibia and humerus of the male C57BL/6-Tg (CAG-eGFP) transgenic mice. 3) Magnetized CD45-eGFP+ cells (donor cells) were obtained after a series of counting, centrifuging, magnetizing and positive sorting. 4) Magnetized donor cells were directly injected into femoral cavities of recipient mice with or without temporary magnetic field (MagIC-TT or non-MagIC-TT group). 5) Survival rate, blood routine examination, the percentage of eGFP+ cells in peripheral blood and pathological examination of the injected femur were analyzed and compared between both groups.Results: 1) Survival curves analysis (Fig.1): at 20th day post-transplantation, the overall survival rate of MagIC-TT group was 72.7% vs 48.5% in non-MagIC-TT group (P=0.083). 2) Blood routine examination: the amount of white blood cells (WBC) in MagIC-TT group was more than non-MagIC-TT group and its P value was 0.03 (Fig.2a). The amounts of hemoglobin (Hb) and platelet (PLT) in both groups weren't shown any significant difference (Fig.2b, Fig.2c). 3) eGFP+ donor cells ratio of peripheral blood (Fig.3): the percentage of eGFP+ cells were gradually increased both MagIC-TT group and non-MagIC-TT group after transplantation, the percentage of the MagIC-TT group seemed to be higher than that of non-MagIC-TT group (P=0.063). 4) Femoral pathological examination: the amount of eGFP+ cells of injected femur in MagIC-TT group was more than those of non-MagIC-TT group at 24 hours post-transplantation.Conclusion: MagIC-TT could enhance donor cells targeted implantation and promote WBC recovery after autologous bone marrow transplantation in C57BL/6 mice. [Display omitted] DisclosuresNo relevant conflicts of interest to declare.

Mini-Review: Regenerating the Corneal Epithelium With Simple Limbal Epithelial Transplantation.

Simple limbal epithelial transplantation (SLET) is an ingenious, low cost and effective technique of limbal stem cell transplantation (LSCT) that is increasingly being undertaken in practice across the world. Since it was first described a decade ago, the technique has been performed in a variety of cases of limbal stem cell deficiency (LSCD) and has underwent several innovative modifications. Published literature on SLET has progressively increased over time and successful outcomes in various clinical scenarios have been reported. This concise review attempts to present a crisp account of SLET covering the indications and contraindications of performing the procedure; detailed account of pre-operative work up and preparation; surgical technique and its modifications; post-operative course, care and possible complications as well as published outcomes of surgery from across the world. Comparative analysis of various techniques of LSCT have been discussed and common concerns of surgeons practising or those who are planning to start practising SLET have been addressed. The authors hope that the pragmatic insights and pearls given at the end of the review will aid the surgeons in performing this technique to provide maximum benefit to patients suffering from the potentially blinding condition of LSCD.