Gamma Irradiation of Human Platelet Lysate:Validation of Efficacy for Pathogen Reduction
and Assessment of Impacts on hPL Performance
Gamma irradiation is one of the most widely employed methods for pathogen reduction andcommercial gamma sterilization facilities areeasily accessible. The whole system for manufacturing gamma irradiated fetal bovine serum(FBS) has been well-established, including doserange, dose mapping, frozen condition, as wellas validation of pathogen reduction. Nevertheless, many research articles have addressed theoptimal conditions for utilizing gamma irradiationin human plasma and blood components. With
these comprehensive references, we previously
assessed the feasibility of using gamma irradiation to obtain pathogen-reduced human platelet
lysate (hPL) and reported low impacts on the potency for cell expansion.

COMPARISON OF PATHOGEN REDUCTION
TREATMENT (PRT) FETAL BOVINE SERUM (FBS)
VS. ULTRAGRO™ GI SERIES VIRAL CLEARANCEVALIDATIONIn this study, we validated the efficacy of gamma
irradiation for virus inactivation. Four model viruses(BVDV, Reo3, HSV1, MMV) were chosen, per ICH/EMA guidelines, to represent a range of viruses
with different genome, structure, size, and sensitivity to various chemical and physical agents. Thevirus spiked hPLs were gamma irradiated and themean values of viral titers showed more than 4log10 reduction across all model viruses. The results demonstrated gamma irradiation is an effective viral reduction procedure for hPL.

25-40 kGy (typicallyemployed for commercialproducts)

To assess the impacts of gamma irradiation on
the long-term stability of hPL performance, weanalyzed UltraGRO™ GI series up to one year after gamma irradiation. The results showed growth
factors still retained comparable levels to the
non-irradiated hPLs. Mesenchymal stromal cells(MSC) cultured with gamma irradiated hPLs formore than three passages showed similar profiles
as with the corresponding non-irradiated hPLs in
respect of growth rate, morphology, immunophenotype, trilineage differentiation potency, and immunosuppressive property.

PRT FBSVs.

UltraGRO™
GI Series

0.22µmSterile filtration

0.22µm<-10º C

Finished productsstorage-20º C

FrozenTransportation toirradiation plant

Frozen on
dry ice

Gamma

Irradiation

Gamma

Cobalt-60

Radiation source

Cobalt-60

Dosage25-40 kGy

Physical state

Sealed bottles

Dry iceTemperature control

Dry ice

FrozenTransportation to
supplier storage

Frozen on
dry ice5-60 kGy (viralinactivation study)

Sealed containers

Virus
CategoryRNA

RNA

DNA

DNA

Enveloped

NonEnveloped

Enveloped

NonEnveloped

Model for

HCV, HIV

HAV

CMV, EBV,
HBV

B19

Virus

BVDV

Reo3

HSV1

MMV

FamilyFlaviReo

HerpesParvo

GenomessRNA

dsRNA

dsDNAssDNA

Size (nm)

40-60

60-80

120-200

18-24

Resistance

Low

Med-High

MediumVery HighUltraGRO™PURE GI> 5.42> 4.40> 4.51

4.55

UltraGRO™
-Advanced GI> 5.54> 4.27> 4.504.46

Reference: AventaCell 2019 ISCT Poster

MSCs Culture with UltraGRO™-PURE GI GMP
Gamma Irradiated/ Viral Inactivate
Xeno-free, Safe, Consistent, Cost effective cell culture

BENEFITS OF ULTRAGROTM PURE GI

AventaCell BioMedical has adopted a state-ofthe-art gamma irradiation process, as a pathogen
reduction treatment (PRT), for viral inactivation tocreate an UltraGRO™-PURE GI (UG-P GI) product.
UG-P GI offers minimized pathogen contamination
risk while preserving potent cell culture performance with human mesenchymal stem cells (hMSCs), human immune cells and other applicable
cell types for clinical applications.

•US FDA DMF # 34284•

JAPAN PMDA Certificate•

Ph. Eur. General Chapter 5.2.12.4 Compliance•

UltraGRO™-PURE GI supplements for producing
clinical grade cells•

Gamma irradiation has been accepted byregulatory agencies as a validated PRT•

Comparable cell culture performance maintained•

Viral inactivated products without loss of potency
Marker %UG-P GI

Cell type

CD73

CD90

CD105

CD34

CD45

CD11bCD79a

HLA-DR

AD-MSC

99.97

99.88

95.33

0.34

0.40

0.78

0.37

1.65

UC-MSC

95.51

99.98

99.09

0.80

0.31

1.08

1.11

1.97

BM-MSC

99.94

99.50

99.95

0.93

0.15

0.15

0.34

1.45

Relative Concentration (%)

Immuno-phenotypical characterization of human MSCs. Human MSCs derived from adipose tissue (AD), umbilical cord matrix (UC), bone marrow (BM) cultured in UltraGRO™-PURE GI for 5 passages displayed characteristic expression of MSC surface markers.

100806040200
EGF

bFGF

IGF-1 PDGF-BB TGF-! 1UG-P

VEGF

UG-P GI

Growth factors retained comparable cytokine levels after receiving
gamma irradiation

MSCs retained immunomodulation potency

Adipo-genesis

AD MSCAD MSC

Adipo-genesis

Osteo-genesis

Chondro-genesis

Human adipose tissue derived MSCs retain tri-lineage differentiation capability after cultured in UltraGRO™-PURE GI supplemented medium for three passages

Osteo-genesis

Chondro-genesis

Reference: AventaCell
UltraGRO™-PURE GI MSC Factsheet

Immune Cell Culture with UltraGRO™-PURE GI GMP
Xeno-free, Gamma Irradiate and Viral Inactivated hPL
for Therapeutic T cell Activation

Ave (n = 5)

1% AB serum

1% UG-P GI

1% plasma

Expansion

1321 folds

2610 folds

1708 folds

T cell %868283Naïve T %

6.0

9.6

2.5

CM T %

5.0

15.5

6

Ave (n = 6)

2% AB serum

2% UG-P GI

2% plasma

Expansion

998 folds

2336 folds

1527 folds

T cell %808578Naïve T %

4.8

7.4

2.5

CM T %

14.4

27.8

8.6

Product Number
HPCHXCGLI05
HPCHXCGLI50

ProductUltraGRO™-PURE GI GMP

Bottle Size (mL)50

PBMCs were collected from 5 healthy donors, and T cells
were activated by applying a commercial kit from supplier A,
followed by the manufacturer’s protocol to compare the induction performance with AB serum, UG-P GI, and auto-plasma. The results showed greater T cells with higher populationof Naïve and central memory T cells could be obtained byintroducing UG-P GI.

PBMCs were collected from
6 healthy donors, and T cells
were activated by applying acommercial kit from supplier B,
followed by the manufacturer’sprotocol to compare theinduction performance with
AB serum, UG-P GI, andauto-plasma. The results
showed greater T cells withhigher population of Naïve andcentral memory T cells could
be obtained by introducing
UG-P GI.

500

Reference: AventaCell UltraGRO-PURE GI
Immune cell Factsheet