Chromosomal aberrations after induced pluripotent stem cells reprogramming

Vaz, Isadora May; Borgonovo, Tamara; Kasai-Brunswick, Tais Hanae; Santos, Dos; Mesquita, Fernanda; Vasques, Juliana Ferreira; Gubert, Fernanda; Rebelatto, Cármen Lúcia Kuniyoshi; Senegaglia, Alexandra Cristina; Brofman, Paulo Roberto Slud

doi:10.1590/1678-4685-gmb-2020-0147

Cited by 8 publications

(6 citation statements)

References 25 publications

(30 reference statements)

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“…At least 50 photomicrographs of metaphase cells were taken for karyotype analysis. Results indicated that the number of chromosomally atypical cells in the hUCMSCs sample was low (Table 2), affirming the genetic stability of hUCMSCs [28].…”

Section: Karyotype Analysis and Identificationmentioning

confidence: 73%

Isolation, Biological Characterization, and Quality Inspection of Human Umbilical Cord-derived Mesenchymal Stem Cells for Clinical Research

Zuo,

Dongcheng,

et al. 2024

AJBGMB

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Background: Human umbilical cord mesenchymal stem cells (hUCMSCs) represent a valuable and versatile cell type derived from the umbilical cord tissue of newborns. These cells exhibit diverse characteristics, including multipotent differentiation potential and low immunogenicity. hUCMSCs hold promise for clinical applications, yet concerns persist regarding their chromosomal stability. This study employs short tandem repeat (STR) analysis to assess genomic integrity. By focusing on chromosomal stability, the study aims to enhance confidence in the clinical viability of hUCMSCs, contributing to their safe and effective use in regenerative medicine. hUCMSCs were extracted from healthy newborn umbilical cords using tissue block adherence and enzyme digestion. The third generation of hUCMSCs undergoes biological characterization and quality checks, including examinations of cell morphology, viability, growth curves, surface markers, cell cycle, and multi-lineage differentiation potential. The result of the present study shows that hUCMSCs exhibit mesenchymal stem cell characteristics, with robust growth and high viability. Positive surface markers are expressed at rates exceeding 95%, while negative markers are ≤2%. Cell karyotyping analysis, STR spectrum identification, and chromosomal microarray analysis confirm genetic stability. Microbial testing shows the absence of contaminants, and immunological studies demonstrate the immunomodulatory capabilities of hUCMSCs. Specific residues, such as trypsin, were not detected. In conclusion, hUCMSCs produced under strict GLP and GMP conditions meet the quality guidelines for clinical application in various stem cell therapies. The findings support the clinical application of hUCMSCs in various medical contexts.

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Section: Karyotype Analysis and Identificationmentioning

confidence: 73%

Isolation, Biological Characterization, and Quality Inspection of Human Umbilical Cord-derived Mesenchymal Stem Cells for Clinical Research

Zuo,

Dongcheng,

et al. 2024

AJBGMB

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“…After reprogramming, hiPSCs regain pluripotency capacity and express proliferation and pluripotency markers like SSEA4, OCT3/4, and Ki67 (Figure 4) 38 , 39 , 40 . It is known that reprogramming, as well as the lengthy culture of hiPSCs and differentiation, can lead to an abnormal karyotype, or rather induce mutations 41 . Therefore, the genetic background of the cultured cells should be monitored over time and at different passages by g-banding and whole exome sequencing.…”

Section: Representative Resultsmentioning

confidence: 99%

Generation of Patient-Derived Podocytes from Skin Biopsies

Rose

Müller-Deile

2023

JoVE

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Podocytes are epithelial cells sitting on the urinary site of the glomerular filtration barrier that contribute to the selective filter function of the glomerulus. Mutations in podocyte-specific genes can cause focal segmental glomerulosclerosis (FSGS), and podocytes are also affected in many other primary and secondary nephropathies.Due to their differentiated nature, primary cell culture models are limited for podocytes. Therefore, commonly conditionally immortalized cells are used. However, these conditionally immortalized podocytes (ciPodocytes) have several limitations: the cells can dedifferentiate in culture, especially when they reach confluency, and several podocyte-specific markers are either only slightly or not expressed at all. This brings the use of ciPodocytes and their applicability for physiological, pathophysiological, and clinical reach into question. Here, we describe a protocol for the generation of human podocytes-including patient-specific podocytes-from a skin punch biopsy by episomal reprogramming of dermal fibroblasts into hiPSCs and subsequent differentiation into podocytes. These podocytes resemble in vivo podocytes much better in terms of morphological characteristics, like the development of foot processes and the expression of the podocyte-specific marker. Finally, yet importantly, these cells maintain patients' mutations, resulting in an improved ex vivo model to study podocyte diseases and potential therapeutic substances in an individualized approach.

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“…Maintenance of genetic stability after reprogramming is required for possible experimental and clinical applications. iPSC lines may show clonal and non-clonal chromosomal aberrations in several passages (from P6 to P34), but these aberrations are more common at of cell cultures in later passages ( Vaz et al, 2021 ).…”

Section: Discussionmentioning

confidence: 99%

Gene-repaired iPS cells as novel approach for patient with osteogenesis imperfecta

Fus-Kujawa,

Mendrek,

Bajdak-Rusinek

et al. 2023

Front. Bioeng. Biotechnol.

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Introduction: The benefits of patient’s specific cell/gene therapy have been reported in relation to numerous genetic related disorders including osteogenesis imperfecta (OI). In osteogenesis imperfecta particularly also a drug therapy based on the administration of bisphosphonates partially helped to ease the symptoms.Methods: In this controlled trial, fibroblasts derived from patient diagnosed with OI type II have been successfully reprogrammed into induced Pluripotent Stem cells (iPSCs) using Yamanaka factors. Those cells were subjected to repair mutations found in the COL1A1 gene using homologous recombination (HR) approach facilitated with star polymer (STAR) as a carrier of the genetic material.Results: Delivery of the correct linear DNA fragment to the osteogenesis imperfecta patient’s cells resulted in the repair of the DNA mutation with an 84% success rate. IPSCs showed 87% viability after STAR treatment and 82% with its polyplex.Discussion: The use of novel polymer Poly[N,N-Dimethylaminoethyl Methacrylate-co-Hydroxyl-Bearing Oligo(Ethylene Glycol) Methacrylate] Arms (P(DMAEMA-co-OEGMA-OH) with star-like structure has been shown as an efficient tool for nucleic acids delivery into cells (Funded by National Science Centre, Contract No. UMO-2020/37/N/NZ2/01125).

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Chromosomal aberrations after induced pluripotent stem cells reprogramming

Cited by 8 publications

References 25 publications

Isolation, Biological Characterization, and Quality Inspection of Human Umbilical Cord-derived Mesenchymal Stem Cells for Clinical Research

Isolation, Biological Characterization, and Quality Inspection of Human Umbilical Cord-derived Mesenchymal Stem Cells for Clinical Research

Generation of Patient-Derived Podocytes from Skin Biopsies

Gene-repaired iPS cells as novel approach for patient with osteogenesis imperfecta

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