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Characterization Services of iPSC Pluripotency

Overview Service Features FAQs Scientific Resources Related Services

Overview

Induced pluripotent stem cells (iPSC) are cells derived from somatic cells with a number of defined factors. They served an important role in cell and tissue modeling, drug screening, and may also in future regenerative therapies. The characteristics of iPSC are similar to embryonic stem cells, thus similar methods are developed to characterize and differentiate them. With years of exploration in iPSC development, Creative Biolabs is dedicated to providing several characterization services of iPSC pluripotency for customers all over the world.

Cell line Characterization and Pluripotency

To characterize the pluripotency of iPSC, a class of defined tests can be used, and usually iPSC are characterized with the criteria applied to ES cells. Due to the differences between iPSC and ES cell expression patterns, their characteristics also not exactly the same. There are a variety of features assessed by characterization, such as cell and colony morphology, the expression of surface markers and other antigens, growth rate, the expression of pluripotency marker genes, enzyme levels, methylation statuses, as well as differentiation into somatic cell types in vitro and in vivo. Besides, among numerous pluripotency markers, OCT3/4 and Nanog are considered to be archetypal pluripotent stem cell markers. Moreover, it has demonstrated that OCT3/4 and SOX2 work together via feedback loops regulating both their own transcription and other pluripotency-associated genes.

Immunofluorescence staining for pluripotency markers. (Lampuoti, J. 2013) Fig 1. Immunofluorescence staining for pluripotency markers. (Lampuoti, J. 2013)

Methods for iPSC Pluripotency Characterization

Creative Biolabs is dedicated to providing several viable and cost-effective characterization methods for iPSC pluripotency. Usually, these approaches are based on RNA extraction, cDNA transcription, polymerase chain reaction, and immunofluorescence staining. Based on our well established iPSC platform, we are confident in offer the first class characterization services to contribute greatly to your project's success. Our services include but are not limited to items.

Markers to distinguish pluripotent and non-pluripotent cells now available for pluripotency marker assays in Creative Biolabs are showing in table 1.

Table 1. Gene and antigen markers analyzed for iPSC pluripotency. (Lampuoti, J. 2013)

Marker Analyzed expression Role/type
c-Myc Gene Expression amplifier, reinforcing the pluripotent state
Nanog Gene, antigen Establishment of induced pluripotency, resisting differentiation
OCT3/4 Gene, antigen Pluripotency maintentance and induction
Rex1 Gene Transcription factor (zinc finger protein), regulated by OCT3/4, SOX2, & Nanog
SOX2 Gene, antigen Pluripotency maintentance and induction
SSEA4 Antigen Stage-specific embryonic antigen, cell surface glycolipid
TRA 1-60 Antigen Stem cell surface keratan sulfate antigen epitope
TRA 1-81 Antigen Stem cell surface keratan sulfate antigen epitope

With professional scientists devoted themselves in iPSC Pluripotency Characterization, Creative Biolabs is dedicated to providing the first class characterization services of iPSC pluripotency for our customers. Please contact us for more information and a detailed quote.

Features of Our Services

Our company's iPSC pluripotency characterization services offer a comprehensive repertoire of features to ensure your iPSCs are properly characterized, ensuring their pluripotency and usability in different research contexts.

  • Advanced technology - Our platform utilizes the latest technologies and methodologies in the stem cell industry. This includes state-of-the-art imaging equipment, cutting-edge molecular biology tools, and sophisticated data analysis programs.
  • Rigorous pluripotency assessment – We conduct stringent tests to confirm pluripotency, including flow cytometry, immunocytochemistry, and differentiation potential assay. We also examine mRNA expression of pluripotency markers to be doubly sure of the cell's capabilities.
  • Differentiation potential verification - To confirm pluripotency, we test the capability of iPSCs to differentiate into cells of all three germ layers (endoderm, mesoderm, and ectoderm) via both in vitro and in vivo procedures.
  • Genomic integrity assessment - Ensuring the genomic stability of iPSCs is crucial. Our services consist of karyotyping, and other genomic tests to check for any mutations or chromosomal aberrations that may have arisen during reprogramming or passaging.
  • Bioinformatics Analysis - We provide an inclusive report that offers a detailed analysis of the test results, interpretation, and possible implications, helping you make an informed decision. Utilizing advanced bioinformatics tools, we establish the 'molecular signature' of characterized iPSCs, essential for their future application in disease modeling, drug discovery, and regenerative medicine.

FAQs

  • Q: What information do I need to provide for you to begin this characterization service?
    A: We would need detailed information about the iPSCs you are sending including their source, the method of reprogramming used, how long they've been cultured, along with any other relevant data you may have collected.
  • Q: How long does the service usually take? And what is the cost of this service?
    A: The duration can vary due to different factors, but generally, we should have a full analysis completed and a report sent to you in approximately 2-3 weeks after receiving your cell sample. The cost varies depending on the type and extent of characterization needed. We can provide you with a custom quote once we have detailed information about your specific requirements.
  • Q: What volume of iPSC confluence is required for service?
    A: The quantity of iPSCs needed can be very dependent on tests required. However, generally, we need approximately 60-80% confluence in a 10 cm dish.
  • Q: What if my samples failed the iPSC pluripotency characterization tests?
    A: If your samples fail the iPSC pluripotency tests, it indicates that the cells are not pluripotent. We can provide you with further guidance on the steps you can take to address this, which could include optimizing your reprogramming protocol or sourcing new iPSCs.
  • Q: Are there any potential risks or challenges involved in iPSC pluripotency characterization?
    A: The primary challenges are usually technical in nature, such as maintaining the optimal growth conditions for iPSCs and potential sample degradation during transport. However, we have established stringent protocols to ensure sample integrity throughout the process.

Scientific Resources

Reference

  1. Lampuoti, J. (2013). “Characterization of Human Induced Pluripotent Stem Cells.”

For Research Use Only. Not For Clinical Use.