NCRAD - The National Cell Repository for Alzheimer's Disease

1. The main function of the National Cell Repository for Alzheimer’s Disease involves collecting, storing and distributing genetic material for Alzheimer’s disease research. Our goal is to provide the scientific community with a valuable resource for obtaining the genetic material needed to study families with Alzheimer’s disease. Researchers from around the world may request DNA or cell lines, the materials used in genetic research. (view image)

2. The process of banking, or storing, DNA and cell lines begins when a blood sample is received from a member of a family having Alzheimer’s disease or serious memory loss.

3. Each sample is assigned a unique kit number. The kit number and information about the participant is recorded in a secured database. (view image)

4. The sample is then taken to the lab and a unique bar code number is assigned to it. The bar code number is entered into a logbook along with the unique kit number. These numbers are checked by several technicians for accuracy. (view image)

5. The blood sample is then placed in a machine and spun to separate the sample into three main layers: the red blood-cell layer, the plasma layer, and the buffy coat, which contains the white-blood cells. The white-blood cells are needed to establish cell lines and obtain DNA.



6a. To establish cell lines, the whiteblood cells are placed in a flask along with a solution that allows permanent cell growth. The cells are incubated at 37°C (body temperature) anywhere from three weeks to three months. (view image)		6b. To isolate DNA, the white-blood cells are washed and spun at a high speed, enabling the cells to cluster together.

7a. The cell-containing solution is then divided and transferred into two larger flasks for further cell growth. It takes approximately one week for the cells to divide to the desired number. The cells are checked throughout this process to ensure they are growing properly.		7b. The cluster of white-blood cells are placed in a solution containing an enzyme that degrades unnecessary cell components. The solution is stored at 37ºC overnight. The cells are split open during this time while the DNA stays intact.

8a. The cells are then placed in a plastic cryo-vial along with a cryopreservative. Each vial holds approximately 1 milliliter of solution containing 1x107 (10,000,000) cells.		8b. Next, the DNA is separated from the cell components. This is accomplished by adding a salt solution and spinning the DNA and cell components at a high speed.

9a. The cells are gradually cooled to freezing temperatures. The slow freeze prevents damage to the cell line and takes place in a controlled- rate freezer.		9b. The DNA is isolated and transferred to another tube, which now contains the desired DNA without the unnecessary cell components. A form of alcohol is added to the solution and interacts with the DNA. This allows the DNA to gather together and become visible. (view image)

10a. The frozen cells are stored in a tank filled with liquid nitrogen at –316° Fahrenheit. Cells can be preserved this way indefinitely and thawed at any time for additional cell growing. This culture and storage process is necessary for immortalizing our participant’s cells for ongoing genetic research in Alzheimer’s disease. (view image)		10b. The DNA is then transferred to a small tube containing a solution that preserves the genetic material for future studies. In a freezer set at –70° Celsius, the DNA is stored for future use. (view image)



11. Qualified researchers around the world continually request samples from the National Cell Repository for Alzheimer’s Disease. A committee reviews each request and determines if the research is appropriate. After a request is approved, the samples are retrieved, thawed, and prepared for shipment. In some cases, a sample is no longer available when all the DNA has been used and the cells no longer grow. If possible, the individual who provided the original sample may be contacted again and asked to provide another blood sample. (view image) With each sample, researchers are given necessary information about the individual’s clinical history and diagnosis. They are never provided with a participant’s identification information, such as their name, in order to protect the confidentiality of the individual and their family. The cell lines and DNA are extremely valuable for Alzheimer’s Disease research. We greatly appreciate all the support from the families who participate in the National Cell Repository for AD.