Human iPS Cell Line (Episomal, CD34+, ApoE3): #iPS26

Product Description

Apolipoprotein (apo) E, a polymorphic protein with 299 amino acids, has important and diverse functions in neurobiology. ApoE distributes lipids among cells in the central nervous system for normal lipid homeostasis and participates in neuronal repair and remodeling. However, the three major human isoforms (apoE2, apoE3, and apoE4) differ in their ability to accomplish these tasks. Human ApoE3 and ApoE4 differ from each another only at one amino acid residue at position 112. ApoE3, the common isoform, has Cys112, whereas ApoE4 has Arg112. ApoE4, the major known genetic risk factor for Alzheimer’s disease (AD), is associated with an earlier onset of AD in a gene dose-dependent manner. It may also contribute to age-related shrinking of the hippocampus and memory deficits in humans. Of note, the lifetime risk estimate of developing AD by age 85 is ~65% in people with two copies of the APOE-ε4 allele, which encodes apoE4, but only ~10% in people with two copies of the APOE-ε3 allele, which encodes ApoE3. This notable difference highlights the importance of ApoE4 in the pathogenesis of AD. Recent studies suggest that apoE4 inhibits hippocampal neurogenesis by impairing neuronal maturation mediated by GABA signaling, ApoE4 increased Aβ production in human neurons leading to GABAergic interneuron degeneration, which could be dramatically ameliorated by treatment with a small-molecule ApoE4-structure corrector. Converting ApoE4 to ApoE3 by gene editing rescued these phenotypes, indicating the specific effects of ApoE4. Neurons that lacked APOE behaved similarly to those expressing ApoE3, and the introduction of ApoE4 expression recapitulated the pathological phenotypes, suggesting a gain of toxic effects from ApoE4. Treatment of ApoE4-expressing neurons with a small-molecule structure corrector ameliorated the detrimental effects, thus showing that correcting the pathogenic conformation of ApoE4 is a viable therapeutic approach for ApoE4-related AD.
ALSTEM’s footprint-free human iPS (induced pluripotent stem) cell line (cat# iPS16) carries APOE-ε4 in both alleles, and was used for generating an isogenic iPSC line (cat# iPS26) with APOE-ε3 in both alleles by changing Arg112 to Cys112. Sequencing results confirmed that this iPSC line has a stable homozygous conversion with Cys112 (TGC) from Arg112 (CGC) in the APOE gene. In addition, another isogenic ApoE deficient (knockout) iPSC line (cat# iPS36) was derived from the same iPS16. The morphology of this human iPS cell line is identical to that of human ES cells. The cells also express the pluripotency markers TRA-1-60, SSEA-3 and Oct4, and demonstrate strong endogenous alkaline phosphatase activity.
ALSTEM’s human iPS (induced pluripotent stem) cell lines (cat# iPS16, iPS26 and iPS36) are isogenic. Lines iPS26 (ApoE3) and iPS36 (ApoE-KO) are derived from line iPS16 (ApoE4). These lines have same genetic background, and are good candidates for studying AD-related pathologies in neurons induced specifically by ApoE4, as well as for drug discovery.
 

Data Analysis


Figure 1. The sequencing results showed this iPSC line (cat# iPS26) had converted Arg112 (CGC) to Cys112 (TGC) with a silence mutation (C to A) in both alleles.
 
 
Product Specifications
Product Name Human iPS Cell Line (Episomal, CD34+, ApoE3)
Catalog # iPS26
Size > 2x105 cells/vial
Shipping Dry Ice
Storage and Stability Store in gas phase of liquid nitrogen immediately upon receipt. This product is stable for 6 months when stored as directed.
Quality Control Human iPS cells were grown in feeder free conditions with mTeSR1 medium. Each lot of human iPS cells is tested for growth and viability following recovery from cryopreservation. In addition, each lot is tested for expression of TRA-1-60 and Oct4, as well as the activity of alkaline phosphatase.
Safety Precaution ALSTEM highly recommends that protective gloves, a lab coat, and a full-face mask always be worn when handling frozen vials. It is important to note that some liquid nitrogen can leak into the vials when submersed in liquid nitrogen. Upon thawing, the liquid nitrogen returns to the gas phase, resulting in excessive pressure within the vial that can cause the vial to explode or expel the cap with dangerous force.
Restricted Use For Research Use Only. Not for use in diagnostic or therapeutic procedures.
Protocol Download
  • Overview
  • Procedure
This protocol can be used for culturing human iPS cells. Footprint-free human iPS cells were generated by transiently introducing episomal plasmids encoding the human transcription factors into human peripheral blood mononuclear cells. The cells were derived using morphological selection criteria and without the use of fluorescent markers or drug selection. When cultured under standard human ES cell culture conditions, the morphology of footprint-free human iPS cells is identical to that of human ES cells. The cells also express the pluripotency markers TRA-1-60 and Oct4, and demonstrate a strong endogenous AP activity.