Karyotype Analysis for iPSC

Creative Biolabs is dedicated to offering the iPSC research and application with the highest-quality karyotyping service. With years of experience, our professional scientists can provide high-efficient iPSC karyotype analysis services for our customers all over the world.

Introduction of Karyotype Analysis for iPSC

The technology of induced pluripotent stem cell (iPSC) lines generation provides a valuable resource for disease modeling, regenerative therapy, and development study. However, the accumulation of karyotypic during long-term iPSCs culture results in chromosomal abnormalities, changes in gene expression and cellular functions, and even increases the risk of the iPSCs being tumorigenic. As genomic alterations present potential risks in the iPSCs applications, it is important to monitor the genomic integrity of these iPSCs lines especially in iPSCs intended for therapeutic use. Now Creative Biolabs has developed various strategies for the iPSC karyotype analysis. iPSC karyotype analysis is the examination of morphology. Moreover, the changes in size, the position of centromeres, and banding patterns are also necessary for detection aspects.

G-bonding Karyotyping for iPSC

The typical strategies for iPSC karyotype analysis are based on staining the chromosomes of metaphase cells in distinct banding patterns. G-banding is a technique to produce a visible karyotype by staining condensed chromosomes. After partially digesting with trypsin, the metaphase chromosomes are stained with Giemsa stain. Heterochromatic regions is more darkly stained in G-banding while less condensed chromatin present light bands in G-banding. We always recommend that a stem cell line should be karyotyped every 10–15 passages to ensure that there are no chromosomal duplications, insertions, deletions, translocations, or centromere loss during the long-term culture. Moreover, we can also provide various karyotyping methods including C-banding (constitutive heterochromatin staining), R-banding (reverse Giemsa staining), T-banding (telomeric staining), and Q-banding (quinacrine staining). In summary, all these methods are based on the abnormality identification according to the changes in banding patterns.

Services at Creative Biolabs

Karyotype analysis is an essential tool in the field of cytogenetics, used to identify and evaluate the chromosomal composition of cells. For iPSCs, maintaining genetic stability is crucial for their use in research and potential therapeutic applications. Our karyotype analysis for iPSC service provides comprehensive chromosomal analysis to ensure the genetic integrity of your iPSC lines.

Our service includes the following key components:

• Sample Preparation
  • We accept iPSC samples cultured and expanded in your laboratory or provide in-house culturing services.
  • Cells are harvested at the appropriate stage to ensure optimal chromosomal spread.
• Chromosome Preparation
  • Cells undergo hypotonic treatment to swell the cells and spread the chromosomes.
  • Fixed in a methanol-acetic acid solution to preserve the chromosomal structure.
• Slide Preparation and Staining
  • Cells undergo hypotonic treatment to swell the cells and spread the chromosomes.
  • Fixed in a methanol-acetic acid solution to preserve the chromosomal structure.
• Microscopy and Imaging
  • High-resolution microscopy is employed to capture detailed images of the chromosomes.
  • Images are analyzed using advanced software to identify and arrange the chromosomes into a karyotype.
• Karyotype Analysis
  • Determining the chromosome number to detect aneuploidies.
  • Identifying structural abnormalities such as translocations, inversions, deletions, and duplications.
  • Preparation of a detailed karyogram for each iPSC line.
• Reporting
  • Comprehensive report detailing the chromosomal status of your iPSC lines, including high-resolution images and descriptions of any abnormalities detected.
  • Optional consultation with our cytogenetics experts to discuss the findings and their implications for your research.

We adhere to stringent quality control measures to ensure the accuracy and reliability of our karyotype analysis. This includes multiple levels of review and validation of results by experienced cytogeneticists. This service ensures that your iPSC lines maintain their genetic integrity, providing confidence in your research outcomes. Let us help you advance your iPSC research with our reliable and comprehensive karyotype analysis service.

Features of Our Service

• Counting: 30-40 cells, and microscope analysis of 20 metaphase spreads.
• A short experimental period of 7 - 10 business days.
• A precise result with high-resolution images.

Fig.1 The karyotype analysis of iPSCs.

Based on our extensive experience in the field of iPSC generation and applications, Creative Biolabs is confident in offering the best services and high-quality products. The deliverables for iPSC karyotype analysis will include a detailed report with high-resolution images to meet your publishing needs. In addition, we can also provide other services regarding iPSC technology, please feel free to contact us if you are interested in them.

Published Data

Below are the findings presented in the article related to karyotype analysis for iPSC.

Lindsay Panther, et al. described a new and reliable non-integrated plasmid reprogramming method for reprogramming fresh peripheral blood mononuclear cells (PBMC) into iPSC (PBMC-iPSC). The iPSCs were subjected to G-band karyotype analysis, which consisted of examining the chromosomes of at least 20 cells in each culture. The results showed that PBMC-iPSC generated using this method had a high rate of karyotypic stability at the chromosome level (Abnormalities in all chromosomes were about 5%, autosomal abnormalities were 2.8%).

Fig. 2 PBMC-iPSCs exhibit stable karyotypes.²

FAQs

• Q: Can you handle large-scale projects involving numerous iPSC samples?
  A: Yes, we are well-equipped to handle large-scale projects. Our lab has the capacity to process multiple samples simultaneously, and we have a dedicated team of specialists to ensure timely and accurate analysis. For large projects, we work closely with you to plan and execute the karyotype analysis efficiently, meeting your research deadlines and objectives.
• Q: What kind of abnormalities can karyotype analysis detect in iPSCs?
  A: Karyotype analysis can detect various chromosomal abnormalities in iPSCs, including aneuploidies (extra or missing chromosomes), translocations (rearrangements of parts between nonhomologous chromosomes), deletions, duplications, and inversions. Detecting these abnormalities is crucial for ensuring the genetic fidelity of iPSCs, which is essential for their reliability in research and therapeutic applications.
• Q: How often should karyotype analysis be performed on iPSC cultures?
  A: It is recommended to perform karyotype analysis on iPSC cultures at regular intervals, typically every 10 to 20 passages, or whenever there is a significant manipulation or stress to the cells. Regular monitoring helps detect any chromosomal changes that may arise during culture, ensuring the ongoing genetic stability of the iPSC lines.
• Q: What are the sample requirements for karyotype analysis of iPSCs?
  A: For karyotype analysis, we require a sufficient number of healthy, actively dividing iPSCs. Typically, a sample of 2-3 million cells is ideal. The cells should be free from contamination and in good condition. We will provide specific instructions for preparing and shipping the samples to ensure their viability upon arrival.
• Q: Can karyotype analysis be used to detect mosaicism in iPSC cultures?
  A: Yes, karyotype analysis can detect mosaicism, which is the presence of two or more genetically distinct cell lines within the same iPSC culture. By examining multiple metaphase spreads from the same culture, we can identify if there is a mixture of normal and abnormal karyotypes, providing insights into the genetic stability and uniformity of your iPSC lines.

Scientific Resources

iPSC Characterization

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References

1. Amit, Michal, et al. "Suspension culture of undifferentiated human embryonic and induced pluripotent stem cells." Stem Cell Reviews and Reports 6 (2010): 248-259.
2. Panther, Lindsay, et al. "Generation of iPSC lines with high cytogenetic stability from peripheral blood mononuclear cells (PBMCs)." BioRxiv (2021): 2021-09.

For Research Use Only. Not For Clinical Use.