Project PXD011783

Summary

Title

Multipotent fetal-derived Cdx2 cells from placenta regenerate the heart

Description

Placental Cdx2 cells can be isolated and utilized for cardiac regeneration in a mouse model of myocardial infarction with evidence of multipotentiality both in vitro and in vivo

Sample Processing Protocol

Pellets were lysed in 100uL of freshly made 8M urea, dissolved in 100mM Tris-­‐HCl (pH 8.5), containing phosphatase and protease inhibitor. Lysates were ultrasonicated to complete lysis and to shear DNA (5 successive 10sec pulses). After determining protein concentrations using a BCA assay, samples were reduced and alkylated (TCEP and fresh IAA), and diluted to 500uL. Samples were digested at 37degrees overnight with constant agitation using Trypsin/LysC mix at a 40:1 ratio. Digests were acidified with TFA and desalted on a 96-­‐well HLB microelution plate and dried to completion prior to MS analysis.

Data Processing Protocol

Samples were resuspended in 0.1% FA at 1ug/uL. 6 uL were injected onto a C18 μPLC column and analyzed under Data Dependent Acquisition mode using an Orbitrap Elite mass spectrometer (ThermoFisher, USA). Average fold­changes values were inferred from the quantified protein­level values as the quotient of the experimental and control groups. These expression differences were subject to ingenuity pathway analysis(IPA).

Contact

Niveda Sundararaman, Cedars Sinai Medical Center
Dr. Jennifer Van Eyk, Cedars Sinai Medical Center, Los Angeles, CA ( lab head )

Submission Date

21/11/2018

Publication Date

09/05/2019

Instrument

LTQ Orbitrap Elite

Software

Not available

Experiment Type

Shotgun proteomics

Publication

    Vadakke-Madathil S, LaRocca G, Raedschelders K, Yoon J, Parker SJ, Tripodi J, Najfeld V, Van Eyk JE, Chaudhry HW. Multipotent fetal-derived Cdx2 cells from placenta regenerate the heart. Proc Natl Acad Sci U S A. 2019 PubMed: 31109997