In this week's video Alex discusses how he finds motivation to keep advocating for research for Parkinson's Disease.
Music: "People Like You" by Weinland
Monday, July 27, 2015
Movie Monday Episode 4 "A New Hope"
In this week's video Alex explains how he raises awareness for Parkinson's.
Music: "Solo Acoustic" by Jason Shaw
Monday, July 20, 2015
Movie Monday Episode 2 "Stem Cell Wars"
In this week's video Alex discusses how Parkinson's has affected his professional and personal life.
Music: "Sunken Eyes" by Weinland
Monday, July 13, 2015
New! Movie Mondays with Alex: Episode 1 "Alex's Menace"
We've gotten a lot of questions about Alex and how Parkinson's Disease has affected his life. To answer some of these questions we will be releasing weekly clips from an interview we conducted with Alex in 2013. The interview answers many of the same questions our fans have asked and will hopefully be insightful, informative, and interesting. In this first clip Alex discusses when he discovered his first symptoms of Parkinson's Disease Enjoy.
Music: "Cold Weather" by Alialujah Choir
Thursday, July 2, 2015
Yes, they can – testing Alex’s cells for pluripotency
In the last blog we gave an overview on how to
characterize iPS cells. Today, we dive in and explain how we
test the cells for pluripotency, the potential to turn into different cell
types of the human body.
Alex: Hi, it's
been a while, but it's summer and I heard
Lauren has accepted a new position!
Birgitt: Yes, we'll miss Lauren and we wish her the best of luck for her future.
Alex: So what's next?
Birgitt: I'll explain how we test your cells to see if they have the potential to
show "yes we can" "Change" into other cells. To test
proteins and markers in the cell you need to visualize them. This can be done
with a method called immunostaining. Immunostaining is an antibody-based method
that detects specific proteins on the surface or inside of the cells. The picture
below is an example of how we can immunostain the cells. We use an antibody
specific for a certain marker on the cell, called epitope. Once this antibody
binds to the epitope we then incubate with a fluorescently labeled antibody
that binds to the first antibody. We can then visualize this fluorescence with
a fluorescent microscope and take the colorful images you will see in the next
few blogs.
More info about immunostaining here
Alex: Hmm sounds a lot like 2008. I posted
some pretty fluorescence pictures on my Facebook page that you shared with me
in the past. I am looking forward to understanding more about these immunostaining
techniques.
Birgitt: We used
three different antibodies to detect whether or not your stem cells are
pluripotent. The three markers we used were OCT4, SOX2, and SSEA4, these are
proteins that are expressed in pluripotent cells and therefore called
pluripotency markers. These are abbreviations for protein names octamer-binding
transcription factor 4 (OCT4), sex
determining region Y-box 2 (SOX2), and stage-specific embryonic 4
(SSEA4). Since these protein names are so complicated we usually only use the
abbreviations.
See the results in the three images below. The blue color is the nuclear stain or
counterstain which shows where the nucleus of the cell is located. The green
fluorescence corresponds one of the pluripotency markers.
Alex: I hope we can talk about this more next time and I am also very excited to come out to San Francisco for the Festival of Genomics on November 5th, 2015 to chat with you on stage about my Parkinson’s disease, stem cells, and more (http://www.festivalofgenomicscalifornia.com/).
Birgitt: I am thrilled to have the opportunity to talk with you on stage, can't wait! And maybe the topic of pluripotency will be reelected in the next post.
 |
iPS colony stained with SSEA-4 antibody |
 |
iPS colony stained with OCT4 antibody |
 |
iPS colony stained with SOX2 antibody |
Birgitt: I am thrilled to have the opportunity to talk with you on stage, can't wait! And maybe the topic of pluripotency will be reelected in the next post.
Wednesday, July 1, 2015
About our New Co-editors
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| Gautam Bordia, Bastian Schuele, Raghav Bordia |
Friday, May 22, 2015
Alex's stem cells pass the practice test
In the last blog we talked about how we bank your cells. Today, we will give you an overview of how to make sure that the cell we have are actually pluripotent stem cells, a process we call characterization.
Lauren: We use four methods to characterize your cells. In the image below you can see a summary of these methods. Two methods test the pluripotency of the newly derived cells and the other two methods make sure that the DNA was not damaged or changed during the reprogramming process.
Alex: I know we have talked about pluripotency before, but can you remind me what that is?
Lauren: Pluripotency refers to the ability of the cells to give rise to all of the cell types that make up the human body. The first way we test pluripotency is by staining the cells for certain markers. All cell types have markers on the surface of their cells and within the cells. These markers are specific to each type of cell so that we can distinguish different cells from one another. We use multiple markers that are specific for pluripotency to be sure that the cells are indeed pluripotent.
Alex: Well that seems fairly straightforward. How else do you test for pluripotency?
Lauren: Another way that we test for pluripotency is to spontaneously differentiate these stem cells. The spontaneous differentiation begins by forming embryoid bodies. Embryoid bodies are 3-D cell aggregates as shown in the picture below. The cells are no longer adherent to the bottom of the dish but are floating in the media. Embryoid bodies are meant to mimic the early development of an embryo. This stage primes the stem cells and gets them ready to differentiate into all cell types. We use specific cell markers to confirm that the stem cells can differentiate into various tissue types.
Alex: I did not realize that so much work went into these processes. What else needs to be done in order to call my newly made stem cells "induced pluripotent stem cells"?
Lauren: The next two methods of characterization ensure that the DNA within the cells is not damaged and that the virus has not "sneaked into" the DNA by the reprogramming process. The first method is called a karyotype which shows the number and shape of chromosomes (packaged DNA) of the cell. For humans, a normal karyotype consists of 22 pairs of chromosomes and the two sex chromosomes X and/or Y. The image below is an example of how a normal karyotype looks like.
Alex: I hope my karyotype comes out normal. What is the next step?
Lauren: To make sure that the genetic material of these reprogrammed cells has not been modified, we measure the virus in the cells which we used to deliver the reprogramming factors. After 10 to 12 passages, we should not detect the virus anymore. We can detect the virus by an amplification method that will be explain in more detail later.
Alex: Thank you for the overview. I am looking forward to learning more about each of these methods and seeing if my cells pass these tests. It is a bit nerve racking almost like passing a practice driving test.
Lauren: Indeed, I have quite nervous about some of the test results of your cells. In the next few blogs we will go into more detail about each of these methods and show the results in you cells. The first process we will discuss is staining for the pluripotency cell markers.
Lauren: We use four methods to characterize your cells. In the image below you can see a summary of these methods. Two methods test the pluripotency of the newly derived cells and the other two methods make sure that the DNA was not damaged or changed during the reprogramming process.
 |
| Summary of characterization process |
Alex: I know we have talked about pluripotency before, but can you remind me what that is?
Lauren: Pluripotency refers to the ability of the cells to give rise to all of the cell types that make up the human body. The first way we test pluripotency is by staining the cells for certain markers. All cell types have markers on the surface of their cells and within the cells. These markers are specific to each type of cell so that we can distinguish different cells from one another. We use multiple markers that are specific for pluripotency to be sure that the cells are indeed pluripotent.
Alex: Well that seems fairly straightforward. How else do you test for pluripotency?
Lauren: Another way that we test for pluripotency is to spontaneously differentiate these stem cells. The spontaneous differentiation begins by forming embryoid bodies. Embryoid bodies are 3-D cell aggregates as shown in the picture below. The cells are no longer adherent to the bottom of the dish but are floating in the media. Embryoid bodies are meant to mimic the early development of an embryo. This stage primes the stem cells and gets them ready to differentiate into all cell types. We use specific cell markers to confirm that the stem cells can differentiate into various tissue types.
 |
| Example of embryoid bodies |
Alex: I did not realize that so much work went into these processes. What else needs to be done in order to call my newly made stem cells "induced pluripotent stem cells"?
Lauren: The next two methods of characterization ensure that the DNA within the cells is not damaged and that the virus has not "sneaked into" the DNA by the reprogramming process. The first method is called a karyotype which shows the number and shape of chromosomes (packaged DNA) of the cell. For humans, a normal karyotype consists of 22 pairs of chromosomes and the two sex chromosomes X and/or Y. The image below is an example of how a normal karyotype looks like.
 |
| Example of a normal karyotype, 46 XY |
Alex: I hope my karyotype comes out normal. What is the next step?
Lauren: To make sure that the genetic material of these reprogrammed cells has not been modified, we measure the virus in the cells which we used to deliver the reprogramming factors. After 10 to 12 passages, we should not detect the virus anymore. We can detect the virus by an amplification method that will be explain in more detail later.
Alex: Thank you for the overview. I am looking forward to learning more about each of these methods and seeing if my cells pass these tests. It is a bit nerve racking almost like passing a practice driving test.
Lauren: Indeed, I have quite nervous about some of the test results of your cells. In the next few blogs we will go into more detail about each of these methods and show the results in you cells. The first process we will discuss is staining for the pluripotency cell markers.
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