Cell Line Creation

Overview

Producer cell lines are the key components for any scientific research and are widely utilized for vaccine manufacturing, antibody production, testing drug metabolism and cytotoxicity, genetics, synthesis of biological compounds, and many more.

Cell Lines can be created from immortal cells such as CHO, Hek293, SF9/SF21, with necessary Vectors to transform them into high producing clones or from primary tissue (healthy and tumor) / primary source (e.g., fibroblast, blood) of human or animal origin. 

Esco Aster provides Cell Line Creation services to ensure the best clone(s) or stem cells can be used for your research, tissue engineering, virus, cell therapy, extracellular vesicles and cellular agriculture.  

We specialize and are best-in-class in accelerated adherent cell line creation and development in conjunction with our adherent cell banking, characterization, testing and storage services. 

Our Process

Esco Aster internally via our collaborator has VERO MCBs/WCBs and Avian cell line from quails with FTO except as used for poxviruses.

We will otherwise create cell lines from global cell banks such as ATCC / ECACC, clients’ RCBs, industrial biotech company cell lines or from academic/research institutes.

our process

OUR SERVICES

Our services are applied in the following areas:

Human Donor Sourcing

Animal Donor Sourcing

Stable Cell Lines (mAbs/Viral Vector)

Allogeneic and Autologous Cell Therapy (MSCs, Fibroblasts)

Immunotherapy

Induced Pluripotent Stem Cells (iPSCs)

Cultivated Meat 

Our Cell Line Creation Workflow


Starting from Tissue Banking Activities

Isolation of Cells

Starting from Cell Source

Modification to Cells
Immortalized Cell Lines from Commercially Available Cell Banks
(e.g. CHO and HEK293)
Gene Editing
Immortalization
Single-cell Cloning
Cell Selection
Research Cell Bank
Differentiation
Induction to Pluripotent Stem Cells
Human Donor Source
Animal Donor Source
Tumor Tissue
Cell Isolation for Cell Therapy
(e.g. TILs)
Blood Source
Cell Sorting
(e.g. Immune cells---T. NK. Dendritic Cells)
Healthy Tissue
Cell Isolation for Stem Cell Therapy
(e.g. MSCs, hESCs)
Primary Cell Isolation for Cell Therapy
(e.g. Fibroblasts, Cardiac, Chondrocytes)
Cell Isolation For Cultivated Meat
(e.g. Adipocytes, Fibroblasts, Chondrocytes)
Primary Cell Isolation for Biologics / Vaccines

Human Donor Sourcing

Upon request and based on donor grid, human donors are screened with optional Clinical Laboratory Improvement Amendments (CLIA) if not equivalent accredited diagnostic laboratories before tissue acquisition. This ensures that primary cells are obtained under an IRB-approved medical ethical consent procedure and signed off by our medical director. 

A variety of stem cells / niche therapeutic cells can be isolated from primary tissue of umbilical cord, cord lining, Wharton’s Jelly, adipose tissue, cardiac cells, fibroblast cells or from bone marrow.  

Depending on the cell type we can develop protocols for enzymatic digestion and utilizing facs to ensure the right cell population is selected or can via tech transfer utilize client protocols. 

For iPSCs we will work to establish required profile of iPSC with client and then identify applicable iPSC lines based on age, gender, disease condition with appropriate reprogramming methodology and matched controls.  Via established 3rd party partners, research institutes, hospitals associated  medical centers and consortiums, we are able to source healthy and patient iPSC lines. 

human donor sourcing

Quality Control Assays

At Esco Aster, all stem cell lines undergo standard quality control assays including USP testing of sterility, mycoplasma testing, pluripotency analysis by fluorescent activated cell sorting (FACS) , and genome integrity by direct polymerase chain reaction (dPCR) and short tandem repeat (STR) analysis. 

quality control

Animal Donor Sourcing

Depending on the species, Esco Aster can aid to source, and with a veterinarian’s help to find an appropriate donor to biopsy for usage in immortal cell line and/or cultivated meat cell line creation. Such animal donors similarly undergo relevant diagnostic testing and screening, as well as relevant institutional review board (IRB) ethics approval to ensure that during a biopsy procedure, no harm was done to animals.

Animal Donor Sourcing

Stable Cell Lines (mAbs/Viral Vectors) 

Esco Aster is able to deliver the expression vectors with the gene of interest into cell lines obtained from human/animal tissue or commercially available cell banks and apply the appropriate cell culture condition with selective pressure to the cells after a short recovery period.

Subsequently, we enrich a high-producing cell population using fluorescence-activated cell sorting (FACS) to improve the efficiency of the potential cell populations for further development in an accelerated timeline.

With full documentation, we implement a state-of-the-art, single-cell dispenser coupled with high-resolution cell imaging which provides image-based evidence of monoclonality that can be used in regulatory filings. By this, we can ensure cell stability and retention of high producer phenotypes over time while achieving stable, high-expressing clones for clinical and commercial manufacturing.

stable cell lines

Cell Therapy (Healthy Tissue Source (MSCs, hESCs, Fibroblasts, etc.)

Mesenchymal Stromal Cells (MSCs) can be isolated from a wide range of tissue sources to be harnessed for autologous or allogeneic therapies. Our isolated MSCs allow faster expansion and greater total expansion numbers due to being younger compared to other commercial sources as we do not over-expand our cells. We expand cells based on [population doubling with characterization to ensure its quality

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cell therapy

Cell Therapy (Tumor Tissue Source)

For immunotherapy, tumor-infiltrating lymphocytes (TILs) can be isolated from a biopsy of tumor tissue and then further isolated for use in further expansion for TIL therapies.

Immunotherapy

Immune Cell Therapy (Blood Source)

Cells may further be selected via antibody bead-based immunoassay or other means to isolate sub-population of cells for usage in autologous immune cell therapy such as in the case of CAR-T, NK Cells.  

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Immunotherapy

Induced Pluripotent Stem Cells (iPSCs)

Cells are either isolated or re-programmed into iPSCs and then banked as master cell banks. For iPSCs, we typically do re-programming and banking from fibroblast, these cells are then differentiated into target cell type for cell therapy or cultivated meat.

The cell biomass is either: 

  • Expanded iPSCs Differentiated in situ Harvest Differentiated Cells 
  • Differentiated In Situ Expand Differentiated Cells Harvest Differentiated Cells

As an option, gene insertion via viral vectors (hyperlink to our viral vector page) or flow electroporation can be used to modify the cell source to either over-express interleukins or for other therapeutic effects. 

Morever, Esco Aster provides stable cell line creation services for viral vector production for usage in genetically-engineered cell therapy or gene therapy. 

Induced Pluripotent Stem Cells (iPSCs)

Cultivated Meat

At Esco Aster, we develop cell sources like animal tissue biopsies and induced pluripotent stem cells (iPSCs) which possess the necessary proliferative capacity and differentiation potential for cultivated meat production.

cultivated meat

Immortalized Cell Lines

We offer immortalized cell lines (e.g., CHO, HEK 293, or GP19) as a host cell from global cell collection centers such as ATCC and ECACC, biotech firms, or research institutes with a comprehensive cell line history document, coupled with chemically defined culture media and appropriate license agreement for further development of manufacturing.

Immoratlized Cell Lines

Gene Engineering

Esco Aster provides clients with proprietary expression vector architecture, codon optimization of gene of interest (GOI), signal peptide screening and high- efficiency transfection system. All stages of research and development processes are with traceable records for the history of host cell line and vector construction.

Gene Engineering

Single-cell Cloning

Esco Aster is implementing advanced single-cell seeding and high-quality imager with full documentation to ensure the monoclonality and to reduce timelines and streamlined process of single cell cloning and developing the integrated strategies for high-throughput clone selection, process optimization and media screening with scale-down model.

Single-cell cloning

Clone Selection

Esco Aster is able to enrich high-producing cell population using fluorescence-activated cell sorting (FACS) and develop the integrated approach for high-throughput clone screening, media and feed optimization using a Design-of-Experiment (DOE) approach in the scale-down model to improve the efficiency of top clone selection in an accelerated timeline.

clone selection

Research Cell Bank

We offer research cell bank (RCB) service for top 3 clones at the end point of cell line creation in GLP laboratory. Meanwhile, we provide clients who have a lead clone they wish to transfer to Esco Aster or who are using Esco Aster’s service platforms of cell line creation for regulatory-compliant master cell bank (MCB) and working cell bank (WCB) services in full GMP facility and appropriately trained and experienced personnel.

Prior to regulatory submission, these cell banks must undergo characterization and biological safety testing to ensure the identity, purity and genetic stability. It ensures the integrity of the product during its development and allows for consistency during manufacturing production.

clone selection

Master/Working Cell Bank

Once the cell line is created this is then banked as master/working cell banks and we work with approved 3rd party vendors for cell line characterization.  

Our cell line developmental services are then provided as next steps in conjunction with culture media development services. 

master/working cell bank
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Workflow