title: Morphology, cell viability, karyotype, expression of surface markers and plasticity of three human primary cell line cultures before and after the cryostorage in LN2 and GN2.
abstract: Primary cell line cultures from human skin biopsies, adipose tissue and tumor tissue are valuable samples for research and therapy. In this regard, their derivation, culture, storage, transport and thawing are important steps to be studied. Towards this end, we wanted to establish the derivation, and identify the culture characteristics and the loss of viability of three human primary cell line cultures (human adult dermal fibroblasts (hADFs), human adult mesenchymal stem cells (hMSCs), and primary culture of tumor cells from lung adenocarcinoma (PCTCLA)). Compared to fresh hADFs, hMSCs and PCTCLA, thawed cells stored in a cryogenic Dewar tanks with liquid nitrogen (LN2), displayed 98.20% ± 0.99, 95.40% ± 1.41 and 93.31% ± 3.83 of cell viability, respectively. Thawed cells stored in a Dry Vapor Shipper container with gas phase (GN2), for 20 days, in addition displayed 4.61% ± 2.78, 3.70% ± 4.09 and 9.13% ± 3.51 of average loss of cells viability, respectively, showing strong correlation between the loss of viability in hADFs and the number of post-freezing days in the Dry Vapor Shipper. No significant changes in morphological characteristics or in the expression of surface markers (being hADFs, hMSCs and PCTCLA characterized by positive markers CD73+; CD90+; CD105+; and negative markers CD14-; CD20-; CD34-; and CD45-; n=2) were found. Chromosome abnormalities in the karyotype were not found. In addition, under the right conditions hMSCs were differentiated into adipogenic, osteogenic and chondrogenic lineages in vitro. In this paper, we have shown the characteristics of three human primary cell line cultures when they are stored in LN2 and GN2.
authors.affiliation: Biobanco del Sistema Sanitario Público de Andalucía (BBSSPA), Centro de Investigaciones Biomédicas, Consejería de Salud - Universidad de Granada, Granada, Spain.
authors.forename: Alberto
authors.initials: A
authors.lastname: Del Pino
authors.affiliation: Biobanco del Sistema Sanitario Público de Andalucía (BBSSPA), Centro de Investigaciones Biomédicas, Consejería de Salud - Universidad de Granada, Granada, Spain.
authors.forename: Gertrudis
authors.initials: G
authors.lastname: Ligero
authors.affiliation: Department of Physiology, Institute of Nutrition and Food Technology, University of Granada, Granada, Spain.
authors.forename: María B
authors.initials: MB
authors.lastname: López
authors.affiliation: Biobanco del Sistema Sanitario Público de Andalucía (BBSSPA), Centro de Investigaciones Biomédicas, Consejería de Salud - Universidad de Granada, Granada, Spain.
authors.forename: Héctor
authors.initials: H
authors.lastname: Navarro
authors.affiliation: Biobanco del Sistema Sanitario Público de Andalucía (BBSSPA), Centro de Investigaciones Biomédicas, Consejería de Salud - Universidad de Granada, Granada, Spain.
authors.forename: Jose A
authors.initials: JA
authors.lastname: Carrillo
authors.affiliation: Biobanco del Sistema Sanitario Público de Andalucía (BBSSPA), Centro de Investigaciones Biomédicas, Consejería de Salud - Universidad de Granada, Granada, Spain.
authors.forename: Siobhan C
authors.initials: SC
authors.lastname: Pantoll
authors.affiliation: Biobanco del Sistema Sanitario Público de Andalucía (BBSSPA), Centro de Investigaciones Biomédicas, Consejería de Salud - Universidad de Granada, Granada, Spain. Electronic address: rafael.diazguardia@juntadeandalucia.es.
authors.forename: Rafael
authors.initials: R
authors.lastname: Díaz de la Guardia
mesh_terms: D000230:Adenocarcinoma; D000077192:Adenocarcinoma of Lung; D050156:Adipogenesis / Q000502:physiology; D000273:Adipose Tissue / Q000166:cytology*; D053687:Adult Stem Cells / Q000166:cytology*; D015703:Antigens, CD / Q000378:metabolism; D015415:Biomarkers / Q000378:metabolism; D002454:Cell Differentiation; D002460:Cell Line; D002470:Cell Survival; D020219:Chondrogenesis / Q000502:physiology; D015925:Cryopreservation / Q000379:methods*; D005347:Fibroblasts / Q000166:cytology*; D006801:Humans; D059785:Karyotype; D007621:Karyotyping; D008175:Lung Neoplasms; D059630:Mesenchymal Stem Cells / Q000166:cytology*; D010012:Osteogenesis / Q000502:physiology; D061251:Primary Cell Culture; D012867:Skin / Q000166:cytology
chemical_list: D015703:Antigens, CD; D015415:Biomarkers
keywords: Dermal fibroblasts; Expression of surface markers; Gas phase of liquid nitrogen; Loss of viability; Mesenchymal stem cells; Primary cell line cultures; Tumour cells