New from REPROCELL:  GMP Grade Stem Cell Master Cell Bank Service Expansion

REPROCELL is pleased to announce the offering of Master Cell Bank (MCB) manufacturing service for the generation of Human Induced Pluripotent Cells (hiPSC) in Beltsville for human therapeutic applications.

REPROCELL has been producing clinically relevant human induced pluripotent stem cells (hiPSCs) by controlling the process from donor recruitment to seed stock hiPSC and encompassing donor eligibility with different geographical regulatory agencies, raw material strict selection, and a proprietary method of reprogramming. REPROCELL has now extended this offering by acquiring a Cytocentric Xvivo system model 2 from BioSpherix, which is a closed GMP cell culturing system. In contrast to conventional cell culture and processing in open incubators and open hoods, the Xvivo System consists of modular sets of closed incubators and closed hoods, integrated as co-chambers and sub-chambers. The enclosed nature of the system allows each chamber to undergo individual environmental control and monitoring. These controls allow greater consistency of the culture environment and reduced risk of contamination, resulting in a superior product and a faster turn-around time. Microscopes, centrifuges, and cryopreservation instruments are also integrated. This technology ensures that we can not only keep our costs low, but also develop the agility to produce a GMP product faster than in a classic GMP room setting. This system provides an ISO 5 quality GMP working environment approved by the FDA. This system will extend REPROCELL's capacity to generate GMP-grade iPSCs master cell banks (MCBs), GMP-grade mesenchymal stem cells, exosomes, and related GMP-grade off-the-shelf cell products for internal use as well as to satisfy the needs of therapeutic, pharmaceutical and biotechnology companies at a competitive price.

REPROCELL has engaged Dark Horse Consulting Group (DHCG), a renowned consulting group specializing in the stem cell sciences arena, to advise in the design and set up of the Cytocentric Xvivo System in our facility. DHCG has developed the User Requirement Specifications (URS) for the implementation of the Cytocentric Xvivo System 2 at REPROCELL. DHCG will be involved in all the steps of developing protocols that will satisfy the US Food and Drug Administration (FDA), the European Medicines Agency (EMA), and the Japanese Pharmaceutical, Devices and Medical Agency (PMDA).

The system:

The Cytocentric Xvivo system model 2 installed at REPROCELL consists of the following modules:

• Module 1 – 3’ Laminar flow Hood
• Module 2 – In-Line Buffer Module
• Module 3 – 57” Sub-Floor Processing Chamber
• Module 4 – In-Line Buffer Module
• Module 5 – 47” Culture Processing Chamber and incubator
• Module 6 – 32” Microscope Module

All modules are completely closed, with aseptic conditions throughout, and advanced controls and monitoring of CO₂, O₂, N₂, relative humidity, and particulate matter. All the modules are physiologically controlled. Figure 1 shows all the modules are installed in a serial fashion providing aseptic conditions, as well as protection from environmental and human contamination of the cells.

The outside environment of the system is controlled by a Clean Room Curtain.  The Clean Room Curtain has two HEPA filters that maintain an ISO-7 environment outside of the Cytocentric Xvivo system as shown in Figure 2.

Module 1 – Laminar flow hood: This hood is used as a staging area to disinfect items going into the system.

Module 2 & 4 – In-Line Buffer Module: Buffer Modules are controlled, double-door, closed environmental chambers that are used as entrance or exit for the Xvivo system or to provide complete isolation from adjacent processing chambers. This ensures that no airborne biological contamination can enter the system. The internal environment of the buffer module is a controlled, cell-optimized atmosphere consisting of CO₂ and  O₂ that can also be ventilated before or after control.

Module 3 – 57” Sub-Floor Culture Processing Chamber: The Culture Processing Chambers (CPCs) are designed specifically to integrate with the incubator module and incubation chamber. Due to their proximity to incubator chambers, these CPCs must be maintained at a maximum aseptic condition. This chamber also has a centrifuge installed in the subfloor. To ensure that CPCs are maintained at maximum aseptic conditions, several control features are utilized. These control features include temperature monitoring, work surface (floor heat) temperature control, relative humidity control, and CO₂ and O₂ control. This CPC and Module 5 are the modules where most of the manual interaction and cell processing occurs.

Module 5 – 47” CPC and incubator: This module includes an incubator. This hypoxia chamber for incubation is integrated into the hoods and opens only into the hood. This eliminates two major shortcomings of conventional open incubators and open hoods. First, incubators can’t become contaminated by personnel or by the environment as they are isolated from both since they open only into the aseptic hood. Second, cells don’t have any disruptions in critical cell parameters because all cell parameters controlled in incubators are also identically controlled in the hood. Additional independent incubation chambers can be fitted into the system for future expansion. Each chamber can be sized for and dedicated to each culture to accommodate any number of different culture protocols simultaneously. Basic cell parameters such as relative humidity, CO₂, and O₂ are fully accounted for, and unique new cell parameters not available elsewhere provide better simulation of physiologic conditions. Every parameter can have dynamic control to adapt to the changing dynamics of any cell population.

Module 6 – 32” Microscope Module: This module is connected to Module 5 through, an inline buffer module that opens into Module 6, which consists of a microscope for visualization of the cells.