Using Blood Cells As a Source for Generating Induced Pluripotent Stem Cells

Sponsored by: STEMCELL Technologies
Session ends: February 27th, 2015, 3:00pm MST
Answers by: Wing Chang, Ph.D., STEMCELL Technologies

Introduction

Human induced pluripotent stem (iPS) cells have the potential to greatly impact many areas of research and medicine and can be generated by reprogramming somatic cells through transient overexpression of key reprogramming factors.

While dermal fibroblasts were the first human cell type to be reprogrammed into iPS cells, blood cells are also increasingly being utilized as a starting cell type due to the limited invasiveness of sample collection, and the availability of banked blood samples representing a variety of disease, age, gender and geographical subtypes.

The choice of cell type to use for reprogramming is based on:

  • The accessibility of tissue samples
  • The genetic make-up of the target cells
  • The reprogramming efficiency


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This session is sponsored by
STEMCELL Technologies

Challenges of Reprogramming Blood Cells

One of the challenges of reprogramming blood cells is the low frequency of certain cell types in peripheral blood. Peripheral blood cell types have varying reprogramming efficiencies, where efficiency is often inversely correlated with frequency in blood. For example, CD34+ hematopoietic stem and progenitors cells have relatively high reprogramming efficiencies, but are rare in circulating blood. In a previous blog post: Reprogramming Blood Cells: How to Decrease Variability in Your Workflow, we discussed some of the solutions that STEMCELL Technologies has developed to isolate and expand rare cell types, such as erythroid and CD34+ progenitor cells, from peripheral blood in order to obtain sufficient numbers for reprogramming.

Another common observed phenomenon is the emergence of partially reprogrammed colonies, usually associated with the continued expression of reprogramming factors. These cells are phenotypically diverse and often fail tests of pluripotency. While overall efficiency is lower, iPS cells that emerged under feeder-free conditions were fully reprogrammed, indicating the importance of culture conditions in the reprogramming process.

Join Wing Chang, Scientist at STEMCELL Technologies in this Ask the Expert Session, as we discuss the technical challenges of reprogramming blood cells, considerations when choosing the somatic cell type to be reprogrammed, implications of starting cell type on reprogramming efficiency and downstream differentiation, and other reprogramming questions.

Don’t miss this chance to have your reprogramming questions answered today!


ask the expert


Questions & Answers

What is your preferred method for confirming pluripotency of iPSCs from blood cells?

Pluripotency can only be confirmed by tri-lineage differentiation either through in vivo formation of a teratoma containing all three germ layers or by in vitro differentiation to cells of each germ layer. Our preferred method would be in vitro directed differentiation using: STEMdiff™ Definitive Endoderm kit for differentiation, STEMdiff™ NIM for neural differentiation, and One […]» Read More

What do you recommend for differentiation of blood cell generated iPSCs – EBs or monolayer?

We typically perform directed differentiation of iPSCs (generated from blood cells) using monolayer cultures. For examples, please refer to our STEMdiff Neural Induction Medium or STEMdiff Definitive Endoderm kits for our monolayer protocols. However, should you decide to differentiate using embryoid bodies (EBs), we would recommend using Aggrewell™ plates for reproducible production of uniformly-sized embryoid […]» Read More

Why would I choose to use blood cells over fibroblasts to reprogram?

Blood cells are continuously produced from stem cells in the bone marrow. The use of blood cells as a source for generating iPS cells is gaining more widespread interest due to the ease in accessibility. Some popular blood cell types used are CD34+ progenitors, erythroid progenitors, T- and B-cells. However, T- and B-cells are less […]» Read More

What are the differences in reprogramming cells on feeder cells vs feeder-free conditions?

Chan et al (1) demonstrated that reprogramming efficiency under feeder-free conditions was lower, but that all of the iPS cells generated were fully (not partially) reprogrammed. Therefore, it seems that culture conditions have an important role in the reprogramming process. That is why we recommend STEMCELL Technologies’ feeder-free, defined, and xeno-free media for reprogramming fibroblasts […]» Read More

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