ASSESSMENT OF MICROBIOLOGICAL CONTAMINATION IN A CELL PROCESSING CENTER FOR A HUMANOID ROBOT○Tomomitsu Iida1,2, Motoki Terada1,2, Yu YuKogawa3, Yumiko Shibata1,2, Michinori Kitagawa1, Shinya Kato1, Tsuyoshi Yorimitsu3, Kenji Matsukuma4, Tadao Maeda1,2, Masayo Takahashi1,2,6, Genki N. Kanda2,4,5,6,○The presenter has no conflict of interest.

1 VCCT Inc. Kobe Eye Center Building 5F, 2-1-8 Minatojima Minamimachi, Chuo-ku, Kobe, Hyogo 650-0047 Japan
2 Kobe City Eye Hospital. 2-1-8 Minatojima Minamimachi, Chuo-ku, Kobe, Hyogo, 650-0047, Japan
3 Technical Research Laboratory Innovation Division, DAI-DAN Co., Ltd. 390 Kitanagai, Miyoshi-cho, Iruma-gun, Saitama 354-0044 Japan
4 Robotic Biology Institute Inc. Telecom Center Building East Wing 1F, 2-5-10 Aomi, Koto-ku, Tokyo 135-0064 Japan
5 Laboratory for Biologically Inspired Computing, RIKEN Center for Biosystems Dynamics Research. 6-2-3 Furuedai, Suita, Osaka 565-0874 Japan
6 Vision Care Inc. Kobe Eye Center Building 5F, 2-1-8 Minatojima Minamimachi, Chuo-ku, Kobe, Hyogo 650-0047 Japan

Background and AimConclusion➢ A robotic cell processing facility (R-CPF), incorporating a versatile humanoid robot system “Maholo”(Robotic Biology Institute Inc.) and an All-in-one cell processing unit (Technical Research Laboratory
Innovation Division, DAI-DAN Co., Ltd), was set up for cell processing for a clinical research.
➢ The air ventilation system with fan filter units kept an atmospheric pressure and the age of air following
the PIC/S-GMP guidelines.

The use of robots in CPF has an issue regarding the impact of robotic structure and operations on anaseptic environment essential for cell processing. In this study, we conducted microbial monitoring toassess cleanliness in R-CPF during cell culture operations by a humanoid robot system “Maholo” for aclinical research of allogenic iPS-derived retinal pigment epithelial cells transplantation.

Method➢ Assessment of microbiological monitoring demonstrated that R-CPF maintained the required cleanliness • Monitoring periods
Microbial monitoring was performed in R-CPF in Kobe Eye Center hospital (Fig.1 and Fig. 2) for a clinicaland an aseptic environment essential for cell processing.
research from August 2021 to June 2022 (Fig. 3). From August to December in 2021, operation
development, operator education and operator training were conducted as Phase 1. From January to May
Discussion
2022, trial operation was conducted as Phase 2. In June 2022, full-scale operation was conducted asPhase 3.
Advantages⚫ Cell culture protocols based on basic science research are directly transfers to a clinical grade cell • Monitoring methods
Microorganisms were monitored in R-CPF as described in Fig.3. All testing locations are shown inprocessing using this system
Fig.3.Floating microorganisms were sampled onto culture plates using an air-sampler at a flow rate of 100⚫ This system reduces labors’ costs for cell culture trainings and cell processing.
⚫ This system reduces involvement of human operators, a primary contaminant, reducing a risk of L/min for 10 min. Falling microorganisms were sampled onto settle plates placed at testing locations for aduration of up to 4 hours. Adhering microorganisms were sampled with sterile swabs or contact plates at
microbial contamination during cell processing.
Limitations
testing locations. All samples were placed in an incubator at 27.5 ℃ for 5 – 7 days. After the incubation,colony-forming units (CFUs) in culture plates were counted. For the swab test, microbial contaminations◼ Culturing multiple types of cells under the current system have a risk of cross contamination.
were determined by either negative or positive of microorganisms in the culture medium.
◼ Several human cell culture skills are difficult to be transferred.

Fig.1, LabDroid Maholo and All-in-one CP unit in Robotic CPF
A, the structural images of RCPF. Left panel shows the 3D
image. A human operator
supports cell culture operationsby LabDroid. Right panel
represents a top view of R-CPF.
Solid arrows indicate flow linesof human. Dot arrows indicate
flow lines of items for cellcultures. B, a top view ofLabDroid and layout of theequipment. The components:① dual-arm humanoid; ② CO2incubator; ③plate carrier slider;④ refrigerator; ⑤ micropipettes;⑥ freezer; ⑦ dustbin; ⑧aspirator; ⑨ tip sensor; ⑩
micropipette tips’ stocker; ⑪
micropipette tips; ⑫workspace;⑬
block
incubator;⑭
microscope; ⑮labware’s stocker⑯centrifuge.
Dural-arm
humanoid performs cell culture
with these peripheral equipment.
C, a front picture of LabDroid.

ABCFig.3, Microbial monitoring results
EFig.2, Cleanliness control and fluid dynamic simulation
A1A2

Robot area
Operator area

Changing roomBC

A, the air circulation system in R-CPF. A1 shows fan filter units (FFUs) on the roof of ーrobot area, ーoperator area, and ーchanging room. A2 shows the air flows in R-CPF as
follows: the flow from outside air to the robot area through the supply-FFUs (straight bluearrow), the flows from the robot to the operator area through an air vent (green arrows), theflows in the operator area purified through the circulation FFUs (circular blue arrows), theflow from the operator area to outside through a damper (red arrows). B, a cleanlinessgrade map. Atmospheric pressures were set for Grade A-C area in R-CPF according toPIC/S-GMP indications. C, Computational fluid dynamics (CFDs) simulation of theairflow inside and outside the R-CPF. CFDs simulation calculates a computation of theage-of-air, a metric denoting the duration necessary for fresh air to replace existing air. Thelowest (3 sec) and highest value (173 sec) of the age-of-air in R-CPF meet the minimum
requirement of the air change rate for PIC/S-GMP indications. Numbers represent the ageof-air (sec). CFDs are simulated at a height of 1,000 mm from the floor.

A–D, positions of microbial monitoring. Box colors: ーGrade A, ーGrade B, ーGrade C. Air sampling, settle plates, swabs, and contact
plates sampling positions are shown in Fig. A, B, C, D, respectively.
Symbols such as ‘A1’ are identical to the position IDs in Fig. E. E,
monitoring results with air sampling, settle plates, swabs, andcontact plates from 2021-09-10 to 2022-06-17. The position IDs are
identical to the symbols in Fig. A–D. The cleanliness classification isbased on the PIC/S-GMP standards. Dark gray, deviation, NT, nottested; INV, invalid (tested but failed). In Phase 1, during operation
development, operator education and operator training, microbial
contamination was detected at various sampling positions in R-CPF.
Besides, the CFUs in September 17, October 8, October 29, andNovember 22 were depart from the accepted standard. In contrast, theacceptable number of microbial colonies were observed at only one
position in one day during trial operation periods, Phase 2. In addition,
no colonies was observed during full-scale operation periods, Phase 3.

Acknowledgement
This study was supported by a grants from AMED and JST-Mirai
Program.