Differentiation of human primary testicular cells in the presence of SCF using the organoid culture system.

PURPOSE: Development of organoids using human primary testicular cells has remained a challenge due to the complexity of the mammalian testicular cytoarchitecture and culture methods. In this study, we generated testicular organoids derived from human primary testicular cells. Then, we evaluated the effect of stem cell factor (SCF) on cell differentiation and apoptosis in the testicular organoid model. METHODS: The testicular cells were harvested from the three brain-dead donors. Human spermatogonial stem cells (SSCs) were characterized using immunocytochemistry (ICC), RT-PCR and flow cytometry. Testicular organoids were generated from primary testicular cells by hanging drop culture method and were cultured in three groups: control group, experimental group 1 (treated FSH and retinoic acid (RA)), and experimental group 2 (treated FSH, RA and SCF), for five weeks. We assessed the expression of SCP3 (Synaptonemal Complex Protein 3) as a meiotic gene, PRM2 (Protamine 2) as a post-meiotic marker and apoptotic genes of Bax (BCL2-Associated X Protein) and Bcl-2 (B-cell lymphoma 2), respectively by using RT-qPCR. In addition, we identified the expression of PRM2 by immunohistochemistry (IHC). RESULTS: Relative expression of SCP3, PRM2 and Bcl-2 were highest in group 2 after five weeks of culture. In contrast, BAX expression level was lower in experimental group 2 in comparison with other groups. IHC analyses indicated the highest expression of PRM2 as a postmeiotic marker in group 2 in comparison to 2D culture and control groups but not find significant differences between experimental group 1 and experimental group 2 groups. Morphological evaluations revealed that organoids are compact spherical structures and in the peripheral region composed of uncharacterized elongated fibroblast-like cells. CONCLUSION: Our findings revealed that the testicular organoid culture system promote the spermatogonial stem cell (SSC) differentiation, especially in presence of SCF. Developed organoids are capable of recapitulating many important properties of a stem cell niche.

Interleukin 4 gene polymorphism (-589C/T) and the risk of asthma: a meta-analysis and met-regression based on 55 studies.

BACKGROUND: Numerous investigations have previously evaluated the association of interleukin (IL) 4 gene polymorphisms and the risk of asthma, conferring inconsistent results. To resolve the incongruent outcomes yielded from different single studies, we conducted the most up-to-date meta-analysis of IL4 gene -589C/T (rs2243250) polymorphism and susceptibility to asthma. METHODS: A systematic literature search was performed in ISI web of science, Scopus, Medline/PubMed databases prior to September 2020, and the pooled odds ratio (OR) and their corresponding 95% CI were calculated to determine the association strength. RESULTS: Literature search led to retrieving of 49 publications (55 case-control studies) containing 9572 cases and 9881 controls. It was revealed that IL4 gene -589C/T polymorphism increased the risk of asthma across all genetic models, including dominant model (OR = 1.22), recessive model (OR = 1.17), allelic model (OR = 1.21), and TT vs. CC model (OR = 1.34), but not the CT vs. TT model. The subgroup analysis by age indicated that IL4 gene -589C/T polymorphism was significantly associated with asthma risk in both pediatrics and adults. Additionally, the subgroup analysis by ethnicity revealed significant association in Asian, American, and Europeans. Finally, subgroup analysis by East Asian and non-East Asian populations indicated significant associations. CONCLUSIONS: The current meta-analysis revealed that IL4 gene -589C/T polymorphism was a susceptibility risk in both pediatrics and adults in the whole and different ethnic groups.

Epidemiologic study of traffic crash mortality among motorcycle users in Iran (2011-2017).

PURPOSE: Motorcycle accident is a major cause of road traffic injuries and the motorcyclists are considered as vulnerable road users. The present study aimed to determine the epidemiological characteristics of fatal motorcycle crashes in Iran. METHODS: In this cross-sectional study, a total of 28,356 motorcycle traffic fatalities registered in the Legal Medicine Organization of Iran were analyzed during the period between March 2011 and March 2017. The examined variables included demographic characteristics, helmet use, crash mechanisms, crash location, position state, type of counterpart vehicle, cause of death and place of death. In the study, road traffic mortalities involving drivers and/or passenger of motorcycles were included. Cases or events registered without these conditions were excluded from the study. To analyse the data, SPSS statistics 25 and GraphPad Prism 8 softwares were used. RESULTS: Of the 122,682 fatal traffic injury cases, 28,356 (23.1%) were motorcycle users, of whom 95.3% were male and 4.7% were female. Most of the motorcycle fatalities belonged to the age group of 18-24 years (29.1%). Head trauma was the major cause of death (59.0%). Also, the overall proportion of safety helmet use among motorcycle crash victims was estimated at 37.4%. Most of the road traffic crash cases (46.8%) happened out of city and half of people (49.9%) died in hospital. About 77.4% of the victims were motorcycle riders and 21.1% were pillion passengers. The highest rate of mortality belonged to the self-employed (38.4%) and then workers (21.8%) and students (10.2%). In addition, most fatalities occurred in people with low education (77.5%) and the least occurred in university graduates (5.5%). Among 31 provinces of Iran, Fars had the highest (9.3%) occurrence rate and Kohgiluyeh and Buyer-Ahmad had the lowest (0.5%). Most of the crash mechanisms were due to motorcycle-vehicle crashes (80.2%), followed by rollover (9.8%). CONCLUSION: Comprehensive public education and special rules are needed to reduce the rate of deaths in motorcycle crashes.

Enhancement and validation of a quantitative GC-MS method for the detection of ∆9-THC and THC-COOH in postmortem blood and urine samples.

Cannabis is frequently detected in forensic investigations and is associated with an increased risk of fatal car crashes. This study aims to develop a method to detect and measure ∆9-tetrahydrocannabinol (∆9-THC) in blood and its metabolite, 11-nor-9-tetrahydrocannabinol-carboxylic acid (∆9-THC-COOH), in urine. The procedure employs two liquid-liquid extraction methods in conjunction with GC-MS in SIM mode. Both compounds were successfully processed, demonstrating the method's ease of use and efficiency. The method was validated for selectivity, identification capability, linearity, precision, limit of detection (LOD), limit of quantification (LOQ), and accuracy. Its effectiveness was further demonstrated by applying it to 30 authentic urine and blood samples from cannabis-related cases, establishing it as a valuable option for routine cannabinoid analysis in forensic toxicology labs.•The linearity range was 25-300 ng/mL for ∆9-THC in blood, and 50-300 ng/mL for ∆9-THC-COOH in urine, and calibration curves for both analytes showed R² values consistently above 0.989, confirming their linearity.•The LOD and LOQ for THC-COOH in hydrolyzed urine were 25 ng/mL and 50 ng/mL, respectively, and for THC in blood, they were 15 ng/mL and 25 ng/mL, respectively.•The variation coefficients were below 14%, and recoveries exceeded 81% for both compounds.

Spatial, geographic, and demographic factors associated with adolescent and youth suicide: a systematic review study.

Background: Suicide is a public health issue and a main cause of mortality among adolescents and the youth worldwide, particularly in developing countries. Objectives: The present research is a systematic review aiming to investigate the spatial, geographical, and demographic factors related to suicide among adolescents and the youth. Methods: In this systematic review, two researchers examined PsycINFO, Web of Science, Scopus, and PubMed databases on December 7th, 2022 with no time limits from the beginning of publication until 2022 to identify the primary studies on spatial and geographic analysis on adolescent and youth suicides. Once duplicate studies were identified and removed, the titles and abstracts of studies were examined and irrelevant studies were also removed. Finally, 22 studies were reviewed based on the inclusion criteria. Results: Our findings show that suicide rates are generally higher among men, residents of rural and less densely populated regions, coastal and mountainous regions, natives, 15-29 age group, less privileged populations with social fragmentation, unemployed, divorced or lonely people, those who live in single parent families, people with mental health issues, and those with low levels of education. Conclusions: Stronger evidence supports the effects of geographic and demographic variables on youth and adolescent suicide rates as compared with spatial variables. These findings suggest that policy makers take spatial and demographic factors into consideration when health systems allocate resources for suicide prevention, and that national policymakers integrate demographic and geographic variables into health service programs. Systematic Review Registration: https://www.crd.york.ac.uk/prospero/, identifier CRD42023430994.

Impact of COVID-19 on poisoning-related mortality in Iran: An interrupted time series study.

BACKGROUND: The COVID-19 pandemic had many negative effects worldwide. These effects involved mental health status issues such as suicide, depression, and the pattern of death associated with drug/poisonings. One of the major concerns of the healthcare community during the pandemic was mortality from poisonings. This study aimed to investigate the trends of mortality from different types of poisonings before and after COVID-19. METHODS: The patients who died from six different categories of drugs or poisons were identified by forensic analysis of body fluids/tissues in Tehran, Iran. The pandemic was separated into the pre-COVID-19 period (April 2018 to January 2020), and the COVID pandemic (February 2020-April 2022). Demographic characteristics were collected from each victim, and comparisons of death trends before and after the pandemic were conducted using the interrupted time series analysis. The absolute number of deaths and proportion of deaths from each type of drug/poisoning were used for the analyses. RESULTS: A total of 6,316 deaths from drugs/poisoning were identified between April 2018-Mar 2022). During this period, 2,485 deaths occurred pre-COVID, and 3,831 were during the COVID-19 era. There were no statistical differences in terms of demographic characteristics before and after the pandemic, except for job status. There was a sharp increase in proportion of methanol death among all poisonings after the start of the pandemic (16.5%, p-value = 0.025), while there was a decreasing trend during the pandemic (-0.915 deaths monthly, p-value = 0.027). The trends for opioids, stimulants, and drug-related deaths changed from decreasing to increasing. No change was seen in the trends for ethanol and volatile substance deaths. This pattern was mirrored in the proportion of each type of poisoning relative to the total number. CONCLUSION: Changes in poisoning-related mortality patterns showed dramatic changes after the start of the pandemic, especially deaths from methanol. Other poisonings such as opioids, stimulants, and drugs should also be addressed as there was an increasing trend during the COVID-19 period, compared to the pre-COVID data.

Preliminary Analysis of Methamphetamine Detection in Lucilia sericata (Diptera: Calliphoridae) Reared in Methamphetamine-Treated Meat at Various Developmental Stages.

Background: It is possible to identify drugs and poisons present in cadavers by analyzing blowfly larvae and pupae collected during forensic autopsies. The main purpose of this study was to use Lucilia sericata's larvae and pupae to identify drugs and poisons present in human cadavers. Methods: In an investigation, immature L. sericata fed meat treated with methamphetamine (MA) at various concentrations (45, 90, and 180 ng/mg) were analyzed to detect MA. Acetylation derivatization and liquid-liquid extraction (LLE) were used as sample preparation methods prior to gas chromatography-mass spectrometry (GC-MS) analytical instrumentation to find MA. Results: According to this study, L. sericata can be used in toxicological testing to identify MA in a host body. All L. sericata larval stages, particularly the third stage larva, pupa and empty pupa tested were positive for MA. Larvae in their first instar produced weak peaks. The L. sericata post-feeding instar following the 45 ng/mg treatment showed the highest MA concentration. For the first time, derivatization using the acetylation approach was used to prepare samples, and successfully, excellent results were obtained. Conclusion: Low quantities of MA can be easily found in immature fly samples using GC-MS. It is important to analyze all samples including human tissues and insect samples, for postmortem drug testing. They can be utilized to find entire MA before they are excreted in excretory samples such as urine. Also, third instar larvae are a great and reliable sample for toxicological study.

VEGF gene polymorphisms in Iranian patients with intracranial glioblastoma.

BACKGROUND: Glioblastoma is one of the most common malignant brain tumors in adults with poor prognosis. Neovascularization is one of the characteristics of these tumors, which is associated with overexpression of vascular endothelial growth factor (VEGF). Accordingly, single nucleotide polymorphisms of this gene could play an important role in structural and functional alterations leading to overexpression of this gene in GBM. METHODS: A total number of 49 patients with GBM and 50 healthy controls were included in the current study. The Genomic DNA was extracted from brain tumor/tissue samples, and after purification assessment, the alleles, and genotypes of rs3025039 and rs2010963 polymorphisms of the VEGF gene were investigated using T-ARMS-PCR. RESULTS: The "T" allele of rs3025039 was 2.79 times more frequent in GBM patients compared to controls (P=0.01). Moreover, the "CT" genotype was 2.83 times more common among patients (P=0.015), while the "CC" was more frequent in controls (P=0.009). The mean overall survival was significantly different between three genotypes of rs3025039, with the longest survival time in "CT" genotype (15.10±5.21, P=0.041). Besides, rs2010963, was significantly associated with GBM occurrence, with the "G" allele being 1.96 times more frequent in patients (P=0.01), as well as the "GG" genotype, which was 7.87 times more common in patients (P<0.001). CONCLUSIONS: Polymorphisms of VEGF could potentially play a role in pathogenesis of GBM, as the allele and genotype distributions of rs3025039 and rs2010963 SNPs were significantly associated with GBM occurrence.

MGMT Gene rs1625649 Polymorphism in Iranian Patients with Brain Glioblastoma: A Case Control Study.

Glioblastoma (GBM) is the most common and aggressive primary malignant brain tumor with poor prognosis and high potential of dispersion to other brain tissues in adult. Effective and modern choices of treatment including chemotherapy with alkylating agents marginally extend survival of GBM. However, alkylating agents can lead to highly harmful mismatch during DNA replication causing apoptosis and cell death. Accordingly, O6-Methylguanine-DNA methyltransferase (MGMT) removes alkyl adducts, thereby causing resistance to alkylating drugs. Single-Nucleotide Polymorphisms (SNPs) in MGMT promoter region may play a role in the regulation of MGMT expression and prediction of glioma development risk. In order to evaluate the clinical significance of rs1625649 SNP in the MGMT promoter region of glioblastoma, genomic DNA from a series of 54 patients with GBM and 50 healthy individuals in Iranian population were collected for tetra ARMS PCR amplification. None of the "A" or "C" alleles were associated with tumor occurrence, the "AA" genotype was more frequent in healthy subjects, and the "AC" genotype was 4.6 times more common in patients with GBM. The longest survival time was observed in the "CC" genotype; however, this difference was not statistically significant. On the other hand, homozygous rs1625649 (AA genotype) was significantly associated with a better survival than the cases with heterozygous rs1625649 (CA genotype) or wild type rs1625649 (CC genotype), predicting better response to temozolomide-based chemotherapy.

Risk of COVID-19 infection and the associated hospitalization, ICU admission and mortality in opioid use disorder: a systematic review and meta-analysis.

BACKGROUND: Opioid use disorder (OUD) as a common drug use disorder can affect public health issues, including the COVID-19 pandemic, in which patients with OUD may have higher risk of infection and severe disease. This systematic review and meta-analysis was conducted to investigate the risk of COVID-19 and the associated hospitalization, intensive care unit (ICU) admission, and mortality in patients with OUD. MATERIALS AND METHODS: A comprehensive systematic search was performed on PubMed, Scopus, Embase, and Web of Science to find studies which compared the infection rate and outcomes of COVID-19 in OUD patients in comparison with the normal population. A random effects meta-analysis model was developed to estimate odd ratios (OR) and 95% confidence interval (CI) between the outcomes of COVID-19 and OUD. RESULTS: Out of 2647 articles identified through the systematic search, eight were included in the systematic review and five in the meta-analysis. Among 73,345,758 participants with a mean age of 57.90 ± 13.4 years, 45.67% were male. The findings suggested no significant statistical relationship between COVID-19 infection and OUD (OR (95% CI): 1.18 (0.47-2.96), p-value: 0.73). Additionally, patients with OUD had higher rate of hospitalization (OR (95% CI) 5.98 (5.02-7.13), p-value<0.01), ICU admission (OR (95% CI): 3.47 (2.24-5.39), p-value<0.01), and mortality by COVID-19) OR (95% CI): 1.52(1.27-1.82), pvalue< 0.01). CONCLUSION: The present findings suggested that OUD is a major risk factor for mortality and the need for hospitalization and ICU admission in patients with COVID-19. It is recommended that policymakers and healthcare providers adopt targeted methods to prevent and manage clinical outcomes and decrease the burden of COVID-19, especially in specific populations such as OUD patients.

Application of Y-STR, DIP-STR and SNP-STR Markers in Interpretation of Forensic Genetic Profiling: A Narrative Review.

Y-STR, DIP-STR, and SNP-STR are useful alternatives for testing the low quantity of DNA in solving the challenges in interpreting forensic genetic profiling. In an unbalanced mixed DNA, partial DNA is often not detected due to the effect of masking by the dominant DNA. Therefore, in such cases interpretation of the results is limited. Furthermore, profiling of these specimens cannot be performed using conventional forensic genetic methods. Biomarkers including Y-STR, DIP-STR and SNP-STR perform well in detecting DNA contributes in the mixed sample. In the present research, the performance of each is evaluated separately.

A trend of Medical Negligence in Laser Therapy in the Capital City: A Nine-Year Survey.

Introduction: The purpose of the present study is to investigate the common causes of injuries, claims, and decisions related to laser therapy medical malpractice during a nine-year survey. Methods: The legal documents in the Coroner's Office of Forensic Medicine were investigated in a national database from 2012 to 2020 in Tehran, Iran. The frequency and nature of the cases, including the year of litigation, the location and certificate of the provider, the injury sustained, and the cause of legal action and judgment were collected. Results: Three hundred and eighty-three cases related to injury from laser therapy were registered in the coroner's Office of Forensic Medicine during the study period. The incidence of litigation related to laser surgery showed an increasing trend, with a peak occurrence in 2020. Laser hair removal was the most common (51.2%) litigated procedure. General practice operators (48%) recorded the highest rate of laser-related medical complaints. Lack of skill was the most common reason for failure. Among 383 cases with public decisions, 62.4% of them were fault liability in paid judgment. Conclusion: Medical claims related to laser application are increasing. However, as it is clear, the growth of laser technology and the increasing demand for lasers in medical science require more surveillance to avoid probable injuries and improve patient safety, especially surveillance of the physicians who work outside the scope of their specialty.

Formation and activity of NLRP3 inflammasome and histopathological changes in the lung of corpses with COVID-19.

COVID-19 is a contagious disease that attacks many organs but the lungs are the main organs affected. The inflammasome activation results in the exacerbation of inflammatory response in infectious disease. The aim of this study is to investigate the formation and activity of the NLRP3 inflammasome complex and the histopathological changes caused by the coronavirus in the lung of deceased persons with COVID-19. In total, 10 corpses; 5 corpses with no history of any infectious diseases and COVID-19 and 5 corpses with the cause of death of COVID-19 were included in this study. Lung tissue samples were harvested during autopsy under safe conditions. Fresh tissues in each group were used to measure the genes expression and proteins level of NLRP3, ASC, Caspase-1, IL-1ß, IL-6 and TNF-α and a routine hematoxylin and eosin staining was performed for histological assessment. Data are represented as the means ± SD. Statistical significance difference was accepted at a p-value less than 5%. The NLRP3, ASC, Caspase-1, IL-1ß, IL-6 and TNF-α genes expression and proteins level were elevated in the lung of the COVID-19 group in comparison with the control group. Histological findings presented the increasing number of polymorphonuclear leukocytes, macrophages and also pulmonary fibrosis in the lungs of corpses with the cause of death of COVID-19. High expression of NLRP3 inflammasome components and its relation with the pathophysiology of the coronavirus-infected lung suggested that targeting the NLRP3 inflammasome could be helpful in achieving a more effective treatment in patients with COVID-19.

Sumatriptan poisoning and its clinical presentation in humans: A case report.

This case report displays some of the possible complications of sumatriptan poisoning, including nephritic syndrome.

An Extracellular Matrix-based Culture System for Generation of Human Pluripotent Stem Cell Derived-hepatocytes.

Liver disease (hepatic disease) adversely affects the normal function of the liver and causes liver problems. Drug-induced liver injury (DILI) can be predicted by primary human hepatocytes. However, the sources of hepatocytes for large-scale drug toxicity screening are limited. To solve this problem, pluripotent stem cells (PSCs), mesenchymal stem cells (MSCs), and hepatic stem cells (HSCs) have emerged as attractive cell sources for cell-based therapies. Human PSCs, including embryonic stem cells (ESCs) and induced pluripotent stem cells (iPSCs) have the ability to undergo self-renewal and differentiate into lineages of ectoderm, mesoderm, and endoderm. Human PSC can be used for the generation of hepatocytes to facilitate the development of novel drugs for the treatment of severe liver diseases. The therapeutic potential of PSC-derived hepatocytes for liver failure have been identified to enhance the development of chemically defined and xenogenic- free 3D culture methods. To date, several hepatic differentiation strategies and various extracellular matrix (ECM) components have been employed to produce hepatocytes or hepatic-like cells (HLCs) in vitro. In this review, we focused on the potential of Matrigel, collagen type 1, Ro- Gel, and laminin as ECM on the differentiation and function of hESC- and hiPSC-derived hepatocytes. The hepatic differentiation of human ESCs and iPSCs would offer an ideal tool for cell therapy and liver diseases.

Validation and Optimization of Ultrasound-Assisted Dispersive Liquid-Liquid Microextraction as a Preparation Method for Detection of Methadone in Saliva with Gas Chromatography-Mass Spectrometry Technique.

Purpose: We investigated validation and optimization of ultrasound-assisted dispersive liquidliquid microextraction (UADLLME) as a preparation method for detection of methadone in saliva samples. Methods: We used blank and methadone-containing saliva samples and also standard methadone solution. Sodium hydroxide and chloroform were added to samples and they were held in ultrasonic bath. Then preparations were centrifuged and extracted analyte was analyzed by gas chromatography-mass spectrometry (GC-MS). Accuracy was measured by Intra and between-day mean relative errors (RE). Precision was assessed by coefficient of variation (CV). Recovery, specificity, linearity and limits of detection and quantification were also determined. Optimization was conducted for ultrasound duration, pH and extraction phase volume. Efficiency of dispersive liquid-liquid microextraction (DLLME) and UADLLME were compared. Results: Intra and between-day accuracies (2.3 -7.5%), recovery (89.4-115.5%) and precision (5.2-11.3%) were all acceptable. Calibration curve was linear in the concentration range of 150 ng/mL-10 µL/mL with R2 >0.9995 and equation of y=86.901x-5342.5. Limits of detection and quantification were 50 and 150 ng/mL, respectively. Specificity was measured by comparing retention times of saliva samples (containing methadone metabolites and other commonly used drugs) during UADLLME/GC-MS analysis and no interference was observed. Recovery of UADLLME was 1.4 of DLLME. Solvent and sample volumes required for UADLLME were 1/200 and 1/20 of DLLME. The greatest efficiency obtained at pH of 10, with ultrasound treatment duration of 5 minutes and extraction phase volume of 1000 µL. Conclusion: Study found that UADLLME/GC-MS is a valid and efficient method for detection of methadone in oral fluid.

Rational MD simulations for improvement the affinity of nanobody against PlGF (placenta growth factor): mutagenesis based on electrostatic interactions.

AbbreviationsCOMcenter of mass distanceMDmolecular dynamicsMM-PBSAMolecular Mechanics Poisson-Boltzmann Surface AreaNbnanobodyPlGFplacenta growth factorRgradius of gyrationRMSDroot mean-square deviationSASAsolvent-accessible surface areaVEGFvascular endothelial growth factor.

The Molecular Mechanism of Aluminum Phosphide poisoning in Cardiovascular Disease: Pathophysiology and Diagnostic Approach.

Nowadays, poisoning with metal phosphides, especially aluminum phosphide (ALP), is one of the main health threats in human societies. Patients suffer from significant complications due to this type of poisoning, and the heart is one of the main organs targeted by ALP. Therefore, in this study, we discussed the effect of phosphine on cardiac function. This study is based on data obtained from PubMed, between 2002 and 2020. The key keywords included "Aluminum phosphide," "Oxidative Stress," "Mitochondria," "Cardiovascular disease," and "Treatment." The results showed that ALP produced reactive oxygen species (ROS) due to mitochondrial dysfunction. ROS production leads to red blood cell hemolysis, decreased ATP production, and induction of apoptosis in cardiomyocytes, which eventually results in cardiovascular disease. Since ALP has the most significant effect on cardiomyocytes, the use of appropriate treatment strategies to restore cell function can increase patients' survival.

Differentiation of human induced pluripotent stem cells into erythroid cells.

During the last years, several strategies have been made to obtain mature erythrocytes or red blood cells (RBC) from the bone marrow or umbilical cord blood (UCB). However, UCB-derived hematopoietic stem cells (HSC) are a limited source and in vitro large-scale expansion of RBC from HSC remains problematic. One promising alternative can be human pluripotent stem cells (PSCs) that provide an unlimited source of cells. Human PSCs, including embryonic stem cells (ESCs) and induced pluripotent stem cells (iPSCs), are self-renewing progenitors that can be differentiated to lineages of ectoderm, mesoderm, and endoderm. Several previous studies have revealed that human ESCs can differentiate into functional oxygen-carrying erythrocytes; however, the ex vivo expansion of human ESC-derived RBC is subjected to ethical concerns. Human iPSCs can be a suitable therapeutic choice for the in vitro/ex vivo manufacture of RBCs. Reprogramming of human somatic cells through the ectopic expression of the transcription factors (OCT4, SOX2, KLF4, c-MYC, LIN28, and NANOG) has provided a new avenue for disease modeling and regenerative medicine. Various techniques have been developed to generate enucleated RBCs from human iPSCs. The in vitro production of human iPSC-derived RBCs can be an alternative treatment option for patients with blood disorders. In this review, we focused on the generation of human iPSC-derived erythrocytes to present an overview of the current status and applications of this field.