1. DUE1104210
STEM CELL TECHNOLOGIES CERTIFICATE PROGRAM
DANIEL PIRSCH
PIRS0001@GMAIL.COM
608-772-0430
EXPERIENCE WITH ADVANCED CULTURE
SYSTEMS, MEDIA AND CELL LINES
 Feeder Dependent – MEF
 Feeder Independent – Matrigel (Geltrex),
Vitronectin (Xeno-Free), Laminin
 mTeSR1, E8, E6, PSC Media, NIM, NDM
 iPSC IMR90-4, hESC H9-WA09
EXPERIENCE WITH ADVANCED CELL
CULTURE TECHNIQUES
 Picking and passaging/maintenance
 Cell Banking – cryopreservation and thaw
 Generation of suspension cell aggregates
 Cell counting and density plating
 Sterile media/reagent formulation
 Tissue culture plate matrix preparation
 Enzymatic and non-enzymatic
lifts/dissociation
ADVANCED CHARACTERIZATION
METHODS
 Photodocumentation
 Morphological characterization and
identification
 Immunoytochemical staining
 G-Banding/Chromosomal Analysis
 Real-Time Quantitative Reverse
Transcription PCR (RTqPCR)
IMR90-4 iPSC’s - MEF/mTeSR-1 Culture
System (400X)
Neural Rosettes (from WA09 H9 hESC’s) on
Laminin (40X)
ICC Image - Neural Precursor Cells stained for
Nestin/DAPI (~400X)

2. EXPERIENCE WITH DIRECTED NEURAL
DIFFERENTIATION METHODS
 7-Day E6 Neuroepithelial Derivation from
hESC’s (Ashton Lab – Wisconsin Institute for
Discovery)
 Accutase single cell dissociation, cell
count and plate at specific density,
culturing in E6 media
EXPERIENCE WITH DIRECTED NEURAL
DIFFERENTIATION METHODS
 Induced Neural Precursor Cell Differentiation –
40+ Days from hESC’s (Zhang Lab – Waisman
Center)
 Cell aggregate generation, suspensions,
plating, maintenance, collagenase IV
dissociation, multiple media formulations
 Demonstrated expression of neural
transcription factors (Nestin, Sox1/2, Pax6,
βIII-Tubulin) by ICC, RTqPCR
EXPERIENCE WITH ENDODERMAL AND
MESODERMAL DIRECTED
DIFFERENTIATION
 Directed differentiation of hESC’s to
Definitive Endoderm and characterization
by RTqPCR
 PSC Cardiomyocyte Differentiation – Life
Technologies Kit
Cardiomyocytes induced from WA09 H9 hESC’s with
PSC Cardiomyocyte Differentiation Kit (40X)
ICC Image – Neural Precursor Cell Stained
for Nestin/Sox2/DAPI (~200X)
Neurosphere in Suspension in Neural Induction
Media (~400X)
OTHER SKILLS/EXPERIENCE:
Laboratory notebook documentation, Biosafety Level 2/WiCell Protocols, Biosafety Cabinet, 37°C/5% CO2
Incubator, mFreSR freezing media, Versene (0.2% EDTA) passaging, manual picking of differentiation,
picking to keep, CHO cell lipofectamine transfection, hemocytometer cell counting, cold transfer matrix prep,
molecular pathways, transcription factors/gene expression, scientific presentations and primary literature
research