New User Special Price Expires in

Let's log you in.

Sign in with Facebook


Don't have a StudySoup account? Create one here!


Create a StudySoup account

Be part of our community, it's free to join!

Sign up with Facebook


Create your account
By creating an account you agree to StudySoup's terms and conditions and privacy policy

Already have a StudySoup account? Login here

PNB 3260 Week 4 notes

by: AnnaCiara

PNB 3260 Week 4 notes 3260

Marketplace > University of Connecticut > Physiology > 3260 > PNB 3260 Week 4 notes
GPA 3.4

Preview These Notes for FREE

Get a free preview of these Notes, just enter your email below.

Unlock Preview
Unlock Preview

Preview these materials now for free

Why put in your email? Get access to more of this material and other relevant free materials for your school

View Preview

About this Document

Lecture 7 and 8
Stem Cell Biology
Dr. Conover
Class Notes
PNB 3260
25 ?




Popular in Stem Cell Biology

Popular in Physiology

This 4 page Class Notes was uploaded by AnnaCiara on Tuesday February 9, 2016. The Class Notes belongs to 3260 at University of Connecticut taught by Dr. Conover in Spring 2016. Since its upload, it has received 14 views. For similar materials see Stem Cell Biology in Physiology at University of Connecticut.

Similar to 3260 at UCONN

Popular in Physiology


Reviews for PNB 3260 Week 4 notes


Report this Material


What is Karma?


Karma is the currency of StudySoup.

You can buy or earn more Karma at anytime and redeem it for class notes, study guides, flashcards, and more!

Date Created: 02/09/16
Week 4 PNB 3260 Lecture Notes Lecture 7 - Student talks on papers Therapeutic Translation of iPSCs for treating neurological disease History  neurogenetic disorders modeled first then sporadic disease Patterns in specific diseases  important for modeling  synaptic defects are different in vivo  Bench and Bedside = working with clinicians Targeted Genome Modification  goal: find similarities between disease phenotype in the lab (dish) and in vivo  Challenge to goal - lower variability  worst to best o ZFNs (zinc-finger nucleases) o TALENs (transcription activator-like effector nucleases) o CRISPR/Cas system - clustered regularly interspaced palindromic repeats  can use more than one mutation if diseases aren't genetic (they are sporadic) harder to model because it's complex lineage specific cells can be used for sporadic cases Neighboring cells  important to consider role of neighboring cells  use other fields to learn about entire environment of the cells Microfluidics  3D growth  allows multiple lines to be grown at once  were able to differentiate half and leave half undifferentiated - ability to choose  helps look at bigger picture: structure and function rather than single cell basis Microelectrode Array  record activity between dif populations of neurons  can be combined with microfluidics  useful for long-term culturing  limitation: accessibility- can't record intracellular potentials iPSC applications  ultimate goal: total regeneration of disease tissue  other goals: o drug development  only 10% make it past safety concerns to be tested  benefits of iPSCs in drug specificity  disease specificity  human specificity  high-throughput screening (HTS) - conduct multiple tests at once o Transplantation therapies  human fetal neural stem cells (fNSCs)  may not work as well as iPSCs  Retinal degeneration  differentiated ESC and iPSC to functional rod photoreceptors  no tumorgenesis in this study  no immunorejection Bench to Bedside  Perfecting cell line's safety and efficacy In Vivo reprogramming in humans  transdifferentiation of cells  fibroblasts and astrocytes from humans o put in 3 growth factors: Ascl!!, Brn2a, Myt1l o became neuronal cells ipscs in clinical and commercial area  commercial o mostly based of unsubstantiated data Stem Cell Reports - PAPER #2 iPSCs to treat ALS in mice  degeneration of upper and lower motor neurons  mostly contralateral but also ipsilateral corticospinal tract mSOD1 mutation  associated with familial  associated with ~20% sporadic cases 2 main hypotheses 1) Glia play a significant enough role in the ALS phenotype that manipulating these cell types will have an observable effect; 2) hiPSCs can be used to generate glial cells that will effectively replace those damaged in ALS, and restore normal glial functionality. hiPSC-GRNPs  glial rich neural progenitors Some mice got hiPSC-GRNPs then littermates got PBS control Male mice had better improvements but died earlier than female mice introduction into in vivo increased amount of neurotrophic factors present mSOD1 induces oxidative stress  males may be more vulnerable to this stress VEGF caused enhanced blood vessel production?  VEGF may be reason for increased life span Study would have had to continue longer to prove it doesn't result in tumorgenicity Lecture 8: Dr. Radmila Filipovic Reprogramming of somatic cells to iPSCs Need autologous cells to avoid host rejection Often fibroblast cells used, hepatocytes also used  use transcription factors  reverse steps of programming to reprogram a cell Zygote is totipotent Pluripotent  multipotent: gives rise to dif cell types in lineage  ICM/ES Unipotent: give rise to only one cell lineage  Stages of development are mimicked in reprogramming zygote-->2 cell-->4 cell-->8 cell-->16 cell-->early blastocyst-->late blastocyst Regrogramming strategies  somatic cell nuclear transfer into eggs  cell fusion -differentiated with undifferentiated  introduction of transcription factors into adult cells using retroviruses  use of transcription factors in viruses that don't integrate into DNA- e.g. adenovirus  small molecules to reprogram  RNA reprogramming???? Generation of iPSCs from MEF cultures via 24 factors  start with mouse embryonic fibroblasts-MEF (fetal human embryonic cells)  introduce transcription factors Test for pluripotency by staining with various biomarkers Factors narrowed down from 24 to 4  Oct4 is rich in the ICM in blastocyst  Yamanaka described these 4 factors Validation of iPS  1) teratoma (type of tumor) formation from ipsc  ability to form different tissues: ectoderm, endoderm, mesoderm, in vivo  3) germline competent adult mice chimera formation after iPS injection in blastocyst o ipsc are giving rise to embryo Kinetics of iPS reprogramming  reprogramming using virally encoded transcription factors takes many days and not all markers of pluripotency are activated together- also low frequency of success  originally 0.05% now up to 2% with retroviral infection  10% with 4 factors + valproic acid (VA) Models  Elite model o predetermined: small number of cells are competent for reprogramming o induced: only cells with specific viral integration sites are competent for reprogramming  Stochastic model: all cells are competent for reprogramming o most supported model Waves of molecular events underlie cellular reprogramming  early phase: mesenchymal to epithelial transition, MET, Cell cycle, decrease cell contact, decrease cell adhesion, microRNA expression, decreased differentiated cell markers, chromatin modifications  intermediate phase: partially reprogrammed cells, resistant cells  late phase: increase in pluripotency markers, DNA methylation, microRNA expression Summary of properties of Oct4, Klf4,Myc, and Sox2  Oct4 critical for making ICM cells pluripotent, active during INTERMEDIATE phase  c-Myc enables self-renewal, active during EARLY phase  Klf4 helps to upregulate Oct4 in embryoid bodies -i.e. supports self renewal, active in EARLY and LATE phase  Sox2 maintains differentiated status, active Will these 4 factors work in humans?  done with the 4 Yamanaka factors  done with variation of the 4 -Thomson: Oct3/4, Sox2, Nanog, LIN28 o worked to generate first line of human iPSCs Other cells that can be reprogrammed  bone marrow, hepatocytes, gastric epithelial cells, pancreatic cells, neural *** Other ways to make iPSCs  because retroviruses integrate into the genome there is interest in finding other ways to make iPSCs  non integrating viruses carrying transcription factors (adenoviruses)  small molecules  direct application of proteins  plasmids (genes cloned in bacteria)  using micro RNAs Direct conversion of fibroblasts to functional neurons by defined factors  done without making ipscs then differentiating them  3 factors: BAM: Brn2, Ascl1, Myt1l  neuron-like morphology in 3 days  form functional synapses  efficiency up to 19% o not all are completely mature o some can fire action potentials with other cell types  narrow 19 factors down to 3 to be important for the reprogramming Modeling schizophrenia using human induced pluripotent stem cells  treatment with antipsychotic drug improves connectivity Advantages of iPSCs over hESCs for preclinical studies  disease specific iPSCs provide renewable source of human cells with genetic background sensitive to disease pathology  one of the reasons why clinical trials failed to examine the effectiveness of drugs is that the effect of a drug maybe different between patients with different underlying mutations  recently iPSC-derived dopaminergic neurons are used to screen group of compounds for neuroprotective Takahashi studies with photoreceptors


Buy Material

Are you sure you want to buy this material for

25 Karma

Buy Material

BOOM! Enjoy Your Free Notes!

We've added these Notes to your profile, click here to view them now.


You're already Subscribed!

Looks like you've already subscribed to StudySoup, you won't need to purchase another subscription to get this material. To access this material simply click 'View Full Document'

Why people love StudySoup

Jim McGreen Ohio University

"Knowing I can count on the Elite Notetaker in my class allows me to focus on what the professor is saying instead of just scribbling notes the whole time and falling behind."

Kyle Maynard Purdue

"When you're taking detailed notes and trying to help everyone else out in the class, it really helps you learn and understand the I made $280 on my first study guide!"

Bentley McCaw University of Florida

"I was shooting for a perfect 4.0 GPA this semester. Having StudySoup as a study aid was critical to helping me achieve my goal...and I nailed it!"


"Their 'Elite Notetakers' are making over $1,200/month in sales by creating high quality content that helps their classmates in a time of need."

Become an Elite Notetaker and start selling your notes online!

Refund Policy


All subscriptions to StudySoup are paid in full at the time of subscribing. To change your credit card information or to cancel your subscription, go to "Edit Settings". All credit card information will be available there. If you should decide to cancel your subscription, it will continue to be valid until the next payment period, as all payments for the current period were made in advance. For special circumstances, please email


StudySoup has more than 1 million course-specific study resources to help students study smarter. If you’re having trouble finding what you’re looking for, our customer support team can help you find what you need! Feel free to contact them here:

Recurring Subscriptions: If you have canceled your recurring subscription on the day of renewal and have not downloaded any documents, you may request a refund by submitting an email to

Satisfaction Guarantee: If you’re not satisfied with your subscription, you can contact us for further help. Contact must be made within 3 business days of your subscription purchase and your refund request will be subject for review.

Please Note: Refunds can never be provided more than 30 days after the initial purchase date regardless of your activity on the site.