Cell Counting

Cellular Dynamics International 

A Fujifilm Company
2

Company Overview

• Cellular Dynamics International (CDI) is the world’s largest 

producer of human iPS cells and iPS cell-derived cell types

• Acquired by FUJIFILM in April of 2015 • Headquartered in Madison, WI , with a site in Novato, CA • Currently employs ~175 total staff • ~750 yrs human stem cell experience • >900 patents (owned or licensed) to enable FTO • Life Sciences and Therapeutics divisions • Core competencies

Creation and culture of human iPS cells

Normal and disease phenotypes

Genetic engineering of iPS cells

SNP repair, Indels, knock-out, knock-in and more

Development of new differentiation protocols

Differentiated cells from all three germ layers

Manufacture of human iPS cell-derived cell types

Scalable production of highly purified cells
April 19, 2016 3

Life Science Research: Current Product Portfolio

iCell 

Cardiomyocytes

iCell Cardiac

Progenitor Cells

iCell 

Neurons

iCell 

DopaNeurons

iCell 

Astrocytes

iCell 

Hepatocytes

iCell 

Hematopoietic 

Progenitor Cells

iCell Endothelial 

Cells

iCell 

Skeletal 

Myoblasts

iCell 

Macrophages

MyCell

Products

Additional 

undisclosed

cell types

Essential 8 Medium

Episomal Reprogramming Kit

Vitronectin

⬤ Quality Management System; Selected guidance 

from both ISO and GMP

⬤ ISO – ideal for research phase

⬤ GMP – required for clinical phase

⬤ 100% complete batch record and traceability; all 

material, incubator, operator, refrigerator, freezer, 

instrument, etc.

⬤ Material management risk profile for every reagent 

on BOM, qualified reagents, 2nd vendors, etc.
4

Cellular Therapeutics: Active Programs

Indication Therapeutic Discovery Pre-clinical IND Phase I

OCULAR

Age-related Macular Degeneration

Retinitis Pigmentosa and Cone 

Dystrophies

CARDIAC

Heart Failure (ischemic cardiomyopathy)

NEURODEGENERATIVE

Parkinson’s Disease
5

Disease Modeling: 250 Donor Panel

Flow Diagram

Blood sample 

collection

Cell Selection 

and expansion Reprogramming Colony picking 

and expansion

iPSC line 

Cryo and QC 

tests

Nuclease-mediated 

genome engineering

Engineered 

line Cryo

Cardiomyocytes 

Cryo and QC 

tests

Selection and 

Expansion

Cardiomyocyte 

differentiation

Cardiomyocyte 

Purification

Cell Count Cell Count Cell Count Cell Count

Cell Count

Cell Count

Cell Count Cell Count Cell Count
April 19, 2016 6

Basis for Data Quality for Cell Counting

QC Control 

Samples

Method 

Qualification

Equipment Qualification IQ

OQ

PQ

Robustness

Accuracy 

Precision

Operator 

Device

Product lot

Product vial

Assay reagent

Daily calibration

QC control sample run 

before each release QC cell 

counting test
April 19, 2016 7

Transition from Cell Counting Device Cedex to ViCell

 Keep using Cedex for existing manufacturing processes 

and develop new processes with ViCell

 Adjust ViCell settings to match cell count numbers from 

Cedex from the same sample

 Adjust ViCell settings to reflect the true cell count, 

understand the cell count differences between Cedex and 

Vicell from the same samples for a certain process step 

and adjust process cell number ranges accordingly
April 19, 2016 8
April 19, 2016 9

Adjust ViCell Settings To Match Cell Count Numbers 

From Cedex

0.0

10.0

20.0

30.0

40.0

50.0

60.0

70.0

80.0

1011009 1011001 1011097 1011147 1011046 1011249

Cells/mL x10

5

Viable Cell Count ViCell

Cedex

D200 D800 H101

0.0

10.0

20.0

30.0

40.0

50.0

60.0

70.0

80.0

1011009 1011001 1011097 1011147 1011046 1011249

Cells/mL x10

5

Total Cell Count 

D200 D800 H101

ViCell Settings Cell Type I

Min Dia 7

Max Dia 50

Cell Brightness 75

Cell Sharpness 40

Viable Cell Spot 

Brightness 80

Viable Cell Spot 

Area 20

Min Circularity 0.3
10

Differentiation Process Example I- Day21 Vicell Setting

0.00

0.20

0.40

0.60

0.80

1.00

1.20

Relative to Cedex

D20/21 Viable Cells/mL

(Relative to Cedex)

P355 in-process NewDA D24-38

Cedex

• Vicell Cell Setting A in-process setting matches the Cedex counts very well (within 8%)

• Vicell Cell Setting A in-process setting matches the reality well enough.

• Recommend this setting for D21 counts
April 19, 2016 CONFIDENTIAL 11

Differentiation Process Example IIImage comparison: 1018075 s1 d11

CEDEX

ViCell
April 19, 2016 CONFIDENTIAL 12

Differentiation Process Example IIImage comparison: 1018136 s1 d11

CEDEX

ViCell
April 19, 2016 CONFIDENTIAL 13

Cell Setting A Cell Setting B

Adjusting Cell Settings for ViCell
April 19, 2016 14

Lesson’s Learned

 Choose cell counting platform wisely to avoid changes: 

Automated 

Meet sample throughput

Reliable and objective results over manual hemacytometer methods Cell 

Record keeping

21 CFR 11 compliance 

Audit trail 

Electronic signature capability 

Secure user sign-on 

User level permissions 

Administrative configuration tools

Easy software update

Good technical and service support

PM
April 19, 2016 CONFIDENTIAL 15

Thank You!