A Short Tandem Repeats (STR) database of human cell lines with 15 loci plus amelogenin

Murcia University and Bioidentity S.L have developed a STR database for comparison and identification purposes, according to the American National Standards Institute (ANSI) recommended protocols: “Authentication of human cell lines: Standardization of STR profiling” ^{[Authentication} of human cell lines: Standardization of STR Profiling. This document is available in: http://webstore.ansi.org/RecordDetail.aspx?sku=ANSI%2fATCC+ASN-0002-2011](ANSI/ATCC ASN-0002-2011). This standard document recommends the next list of 8 core STR markers, plus amelogenin, to uniquely identify human cells:

The standard also establishes a set of match criteria for human cell line authentication, based on an algorithm, which allows to compare the number of shared alleles between two cell lines and to express it as a percentage:

• Cell lines with ≥ 80% match are considered to be related; derived from common ancestry.

• Cell lines with a percent match of 56-79% are probably unrelated, but this range includes a small number of related samples, and may need further testing.

• Unrelated cell lines generally show percent match figures of 55% or less.

ANS-0002 standard document concludes that “98% of related cell line samples can be successfully authenticated using the matching algorithm, with ≥ 80% match as the threshold”. However, the document also admits that further data, such as analysis of additional STR loci, can be required to discriminate among certain samples which give results in the 56-79% match range. International Cell Line Authentication Committee (ICLAC) shares this opinion (ICLAC Match Criteria Worhsheet v 1.1). Moreover, in some cases, the addition of more markers will allow to discriminate between closely related cell lines, which without an increased number of loci can be established as derived from same ancestry but not differentiated further away.

Lastly, our experience suggests ≥ 80% match is a valid threshold that two related lines almost always must observe, although unrelated ones can sometimes reach it as well. Therefore, we prefer to write in our authentication reports: “a result with ≥ 80% match is consistent with two samples being related” (wording from ICLAC Match Criteria Worhsheet v 1.1) instead of “cell lines with ≥ 80% match are considered to be related” (more emphatic wording from ANS-0002 standard document).

For this all reason, we decided to analyze, and include in our database, 7 more loci than the 8 core STR markers in order to clarify doubtful cases of human cell lines authentication, since main online interactive database (ATCC, DSMZ) do not normally give this information. To this end, we use the AmpFlSTR® Identifiler® Plus PCR Amplification Kit (Applied Biosystems) that amplifies 15 STR loci and the amelogenin gender-determining marker. Besides the 9 recommended markers by ANS-0002 document, the 7 additional loci are:

Our criterion to establish reference data for 16 loci comparison

Research Support Services of Murcia University and Bioidentity S.L. mainly operate as supplier of STR profiles and human cell line authentication analysis (Aquí, un enlace a nuestros servicios de autentificación), always working according to the ANS-0002 standard document. After obtaining 8 STR + AMEL profile for a sample, it can be considered to be authenticated when it complies with the match criteria of the ANS-0002 document. However, a cell line authenticated in our lab will only reaches the quality of reference data for 16 loci comparison purposes, when it exactly shows all the same alleles (for the 8 core STR markers + AMEL) as its presumed identity dictated. Therefore, our criterion calls for 100% identity in type and number of alleles. When it is achieved, the whole profile (15 STR + AMEL) will be considered as a valid reference data for comparison in our database.

In our database, the search options include 8 or 15 loci + AMEL. The search results shows % Match resulting from comparison between user data and our resource for human cell line STR profiles (references). The algorithm and match criteria recommended by ANS-0002 document are used.

Bioidentity

Bioidentity is a specialized company in genetic analyzes to determine the genetic profile in cells or individuals.

DNA has differents markers, the genetic fingerprinting can be determined when they are analiced, curretly this is the most reliable and precise method of individual identification, it is increasingly common to find new applications and utilities to this valuable information.

Our mission is to provide genetic tools to any person, company or institution to identify individuals or resources used in scientific research. We prtend to offer a specialized and great value to our customers.

Our laboratory is located at the Business Innovation Centre for Biotechnology (BSIC) of Miguel Hernández University of Elche, that place is an innovative and high technological level environment. We have established scientific links and professional collaboration with that university, spreading to other important institution. These relationships have helped to Bioidentity to grow as a company of future in the biotechnological field.

Location

Parque científico empresarial, Miguel Hernández University
Edificio Quórum III
Avenida Universidad s/n 03202, Elche, Alicante, Spain
www.parquecientificoumh.es/cieb

Contact

Email: contacto@bioidentitysl.com
Phone: +34 603 85 90 62

Sección de Apoyo Estadístico, Servicio de Apoyo a la Investigación

Apoyo Estadístico (AE) ofrece a la Comunidad Universitaria, así como a entidades públicas y privadas, la aplicación de la Estadística, que junto con los medios informáticos actuales, nos permite analizar y corroborar con la realidad, los fenómenos experimentales objetos de estudio.

Location

Edificio SACE (n. 21)
Campus de Espinardo, University of Murcia
30100 Murcia, Spain

Contact

Email: amaurandi@um.es, sai.sae@um.es
Phone: 868 88 7315
Fax: 86 888 7302

Sección de Biología Molecular, Servicio de Apoyo a la Investigación

Facilitar la puesta a punto y mantener aquellas técnicas de biología molecular, que requieran los departamentos de la Universidad de Murcia para la realización de las labores de investigación propias de la actividad universitaria.

Location

Edificio CAID (n. 21)
Campus de Espinardo, University of Murcia
30100 Murcia, Spain

Contact

Email: sai.sbm@um.es
Phone: 868 88 8187
Fax: 86 888 7302

Sección de Cultivo de Tejidos, Servicio de Apoyo a la Investigación

Su función consiste en facilitar la puesta a punto y mantener aquellas tecnologías de cultivo de células que requieran sus usuarios, para la realización de las labores de investigación propias de la actividad universitaria.

Location

Edificio SACE (n. 21), Floor 1
Campus de Espinardo, University of Murcia
30100 Murcia, Spain

Edificio LAIB, Floor 0
Campus Ciencias de la Salud, University of Murcia

Contact

Email: sai.sct@um.es
Phone: 868 88 7157
Fax: 86 888 7302