Multipotent fetal-derived Cdx2 cells from placenta regenerate the heart
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).
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