The Avon Longitudinal Study of Parents and Children (Alspac), popularly known as the Children of the 90s study, has built a reputation as a unique, ongoing research project
A pioneering study of child health based at the University of Bristol is about to enter a new phase, thanks to a new automated cell line system developed with RTS.
The Avon Longitudinal Study of Parents and Children (Alspac), popularly known as the Children of the 90s study, has built a reputation as a unique, ongoing research project.
14,000 mothers were enrolled during pregnancy in 1991-2 and both the children and their parents have been followed in minute detail ever since.
By studying both the children's genes and their environment, Alspac hopes to shed more light on the causes of diseases such as heart disease, allergies, diabetes, attention deficit disorder, depression, and the autistic spectrum disorders.
Over the last 12 years, data has been amassed from questionnaires, direct tests and examinations of the children, linked to medical and educational records.
Half a million biological samples have also been taken - everything from placentas to milk teeth.
With assistance from the Wellcome Trust, Alspac is setting up a DNA bank in new laboratories built in the shell of the old Bristol Children's Hospital.
RTS Life Science International has created a system to manage and carry out the cell manipulations crucial to this project.
Richard Wynn Jones, head of biological samples in Alspac, explained: "It is unusual to automate a process as variable as this, but it is also unreasonable to ask people to perform such large numbers of repetitive tasks.
"We were also worried about the accuracy of our data and the possibility of contamination and infection.
"RTS has created a modular system that will allow us to automate other parts of our process in the future, while still being controlled by the same scheduling software. "Essentially the system automates the tissue culture process that is vital to the development of immortalised cell lines, which will provide a never ending supply of DNA for extensive genetic studies.
"We have a target of 25,000 cell lines to be created over five years".
The taking of blood samples is an important part of the study's ongoing analysis, and these are used for a variety of purposes, including the generation of cell-lines.
Peripheral blood lymphocytes in blood are isolated and transferred to a single well of a 24 well plate in which they are exposed to Epstein Barr Virus (EBV).
This results in a single stock well.
Once set up, the transforming cells are subjected to the automated refeeding and expansion process.
Cells are diluted and expanded progressively across the wells of the plate.
Plates are barcoded and strictly audited throughout their entire expansion process.
The barcode on the plate is read by the system and the appropriate refeeding or expansion protocol selected. Either a proportion of the medium is removed and replaced, or the cells are diluted with medium and expanded into a larger number of wells.
The plate is returned by the system to its incubator prior to a further inspection after several days.
The growing cells move through the system until all 24 wells contain confluent cells.
The cells are then transferred to flasks for a final expansion phase before being aliquoted for cryostorage and DNA extraction.
The RTS system at Bristol University is capable of handling up to 300 cultures at any one time, with inspections taking place during office hours and expansion and cell feeding taking place overnight.
On average, there are 1200 manipulations each week.
If the system runs low on consumables, such as the liquid handling tips, Sprint remotely messages the lab staff.
If there is no response, or it is instructed to do so, it will finish the part-processed plates and return them to the incubators overnight, ensuring no valuable cells are left stranded by the system.
RTS has employed the same principles to the Bristol University system as the company applies to its assay platform systems for high throughput and ultra high throughput screening.
Automated assessment and expansion of cell lines is already provided on other RTS cell culturing systems.
David Harding, RTS business development manager explained: "The success of assay platform, and the flexibility inherent in both the physical layout and the Sprint scheduling software it uses, translates very well to producing a fully automated system for cell manipulation in multi-well plates." This system type has the capacity for automated plating out, re-feeding, well assessment (via automated 'visual' assessment or Elisa assay) and cherry picking of wells for expansion.
Enclosing the system within a class II environment means that a range of applications such as Hybridoma production, vaccine development, DNA banking, Caco2 plate production and assaying are, for the first time, served in a fully automated manner.