Optimizing DNA Yield From Buccal Swabs in the Elderly: Attempts to Promote Buccal Cell Growth in Culture DAVID J. VANDENBERGH, 1,2, * KATE ANTHONY, 1 AND KEITH E. WHITFIELD 2 1 Center for Developmental and Health Genetics, The Pennsylvania State University, University Park, Pennsylvania 16802 2 Department of Biobehavioral Health, The Pennsylvania State University, University Park, Pennsylvania 16802 ABSTRACT Participation in genetic studies is often limited by a volunteer’s reluctance to donate blood samples. We wished to determine if alternate, less painful, methods to venipuncture could be used to collect cells to provide DNA for genotyping, and whether the cells could be grown in culture for extraction of DNA. Volunteers in the study were comprised of two groups. Nine individuals from a university campus were recruited to provide samples for initial experiments. A second group of 710 twins and singletons from North Carolina and of African-American descent were a part of an ongoing study of age-related traits and participated in collection of buccal swabs via the mail. A protocol was generated that maximizes the recovery of DNA from buccal swabs, which are easier to handle than saline rinses. The DNA recovered is stable over several years, allowing genotype tests at a future date. Attempts to encourage growth of buccal epithelial cells recovered from swabs in tissue culture proved unsuccessful. Buccal swabs work well for the collection of DNA, especially from nonclinic-based volunteers, and can be sent via the mail to the laboratory for DNA extraction. Thus, an inexpensive and efficient method exists for genetic studies of population-based samples. Am. J. Hum. Biol. 15:637–642, 2003. # 2003 Wiley-Liss, Inc. For many genetic studies, recruiting suf- ficient numbers of volunteers has been hampered by the necessity of drawing blood to recover sufficient DNA for genotype analysis. Not surprisingly, most people will avoid the pain involved in this procedure if possible. This reluctance may vary from population to population, depending on their cultural differences. Greater care is also required with blood collection than with other types of samples due to the risk of infection for the volunteer and the researcher. The result is less available data on the genetics of understudied populations who may not be able, or willing, to travel to centralized blood collection facilities. Given the general aging of American populations, and that African-American elders represent one of the fastest-growing segments of this population (Angel and Hogan, 1992; Himes, 1992), fewer data will be collected on these important groups. Successful recruitment of African- Americans to biomedical research studies, whether clinic-based or more epidemi- ological, presents additional difficulties. In part, this difficulty is due to previous unethical and poorly designed research, including the Tuskegee Experiment (Bowman, 1991; Gamble, 1993). Another concern expressed by many potential African-American sub- jects is the possibility that the risks asso- ciated with what are deemed to be well- designed and ethical research might not be linked to improved health of the community being studied (King, 1992). The ability to overcome these challenges rests on multiple approaches (Whitfield, 2001), one of which is to make the research protocols as simple, convenient, and harmless as possible— especially genetics-based biomedical research. The development of polymerase chain reaction (PCR) allows for genetics studies using only nanogram quantities of DNA, obviating the need to collect large amounts of DNA, such as from blood samples. The success in producing genotypes with small quantities of DNA has been partially offset ß 2003 Wiley-Liss, Inc. Contract grant sponsors: the College of Health and Human Development, The Pennsylvania State University, CAATSA is funded by a grant from the National Institute on Aging; Contract grant number: 1RO1-AG13662-01A2 (to KEW). The author have no affiliations or financial interests that would influence the subject matter discussed. *Correspondence to: D.J. Vandenbergh, Department of Biobehavioral Health, The Pennsylvania State University, 101 Amy Gardner House, University Park, Pennsylvania 16802. E-mail: djv4@psu.edu Received 23 September 2002; Accepted 7 March 2003 Published online in Wiley InterScience (www.interscience. wiley.com). DOI: 10.1002/ajhb.10177 AMERICAN JOURNAL OF HUMAN BIOLOGY 15:637–642 (2003)