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Establishment and Preservation of Lymphoblastoid Cell Lines from Fresh and Frozen Whole Blood and Mononuclear Cells.
Asadi, Masoumeh; Ganjibakhsh, Meysam; Aghdam, Samaneh Mahmoud; Izadpanah, Mehrnaz; Moghanjoghi, Shiva Mohamadi; Gorji, Zahra Elyasi; Rahmati, Hedieh; Amoli, Abdolreza Daneshvar; Movassagh, Sepideh Ashouri; Fazeli, Seyed Abolhassan Shahzadeh; Farhangniya, Mansoureh; Farzaneh, Parvaneh.
Afiliação
  • Asadi M; Human and Animal Cell Bank, Iranian Biological Resource Center (IBRC), ACECR, Tehran, Iran.
  • Ganjibakhsh M; Human and Animal Cell Bank, Iranian Biological Resource Center (IBRC), ACECR, Tehran, Iran.
  • Aghdam SM; Human and Animal Cell Bank, Iranian Biological Resource Center (IBRC), ACECR, Tehran, Iran.
  • Izadpanah M; Human and Animal Cell Bank, Iranian Biological Resource Center (IBRC), ACECR, Tehran, Iran.
  • Moghanjoghi SM; Human and Animal Cell Bank, Iranian Biological Resource Center (IBRC), ACECR, Tehran, Iran.
  • Gorji ZE; Department of Genetics, Faculty of Biological Sciences, Tarbiat Modares University, Tehran, Iran.
  • Rahmati H; Human and Animal Cell Bank, Iranian Biological Resource Center (IBRC), ACECR, Tehran, Iran.
  • Amoli AD; Human and Animal Cell Bank, Iranian Biological Resource Center (IBRC), ACECR, Tehran, Iran.
  • Movassagh SA; Human and Animal Cell Bank, Iranian Biological Resource Center (IBRC), ACECR, Tehran, Iran.
  • Fazeli SAS; Human and Animal Cell Bank, Iranian Biological Resource Center (IBRC), ACECR, Tehran, Iran.
  • Farhangniya M; Human and Animal Cell Bank, Iranian Biological Resource Center (IBRC), ACECR, Tehran, Iran.
  • Farzaneh P; Department of Molecular and Cellular Biology, Faculty of Basic Sciences and Advanced Technologies in Biology, University of Science and Culture, Tehran, Iran.
In Vitro Cell Dev Biol Anim ; 56(4): 332-340, 2020 Apr.
Article em En | MEDLINE | ID: mdl-32358742
ABSTRACT
Although blood cells are interesting sources for genome investigations, one of the main problems in obtaining genomic DNA from blood is the restricted amount of DNA. This obstacle can be avoided by generating Epstein-Barr virus (EBV)-induced B cell lines. This study investigates the efficiency of four different methods to generate lymphoblastoid cell lines (LCLs). Blood samples (n = 120) were obtained from donors and categorized into four groups fresh whole blood, frozen whole blood, fresh peripheral blood mononuclear cells (PBMCs), and frozen PBMCs. The samples were followed by EBV transformation to generate LCLs. Quality control and authentication of the cells were performed using multiplex PCR and short tandem repeat (STR) analyses. Finally, we assessed the success rate and amount of time to establish the cell lines in each group. The results showed that the cells were not contaminated nor were they misidentified or cross-contaminated with other cells. The success rate of LCLs generated from the whole blood groups was lower than the PBMC groups. The freezing procedures did not have any considerable effect on the establishment of lymphoblastoid cells. These established cells have been preserved in the human and animal cell bank of the Iranian Biological Resource Center (IBRC) and are available for researchers. Due to the management and transformation of a substantial number of blood samples, we recommend that researchers freeze PBMCs for further use with high efficiency and time-saving. We suggest that whole fresh blood should be directly transformed when the volume of the blood sample is less than 0.5 ml.
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Texto completo: 1 Base de dados: MEDLINE Assunto principal: Preservação Biológica / Leucócitos Mononucleares / Linfócitos / Técnicas de Cultura de Células / Congelamento Limite: Adult / Humans / Middle aged Idioma: En Revista: In Vitro Cell Dev Biol Anim Assunto da revista: BIOLOGIA Ano de publicação: 2020 Tipo de documento: Article País de afiliação: Irã

Texto completo: 1 Base de dados: MEDLINE Assunto principal: Preservação Biológica / Leucócitos Mononucleares / Linfócitos / Técnicas de Cultura de Células / Congelamento Limite: Adult / Humans / Middle aged Idioma: En Revista: In Vitro Cell Dev Biol Anim Assunto da revista: BIOLOGIA Ano de publicação: 2020 Tipo de documento: Article País de afiliação: Irã