Tags

Type your tag names separated by a space and hit enter

Hair follicle-derived mesenchymal cells support undifferentiated growth of embryonic stem cells.
Exp Ther Med. 2017 May; 13(5):1779-1788.ET

Abstract

The aim of the present study was to investigate whether feeder layers composed of human hair follicle-derived mesenchymal stem cells (hHFDCs) are able to support human embryonic stem cells (hESCs). hHFDCs and mouse embryonic fibroblasts (MEFs) were isolated and cultured in Dulbecco's modified Eagle's medium (DMEM)/F-12 and low-glucose DMEM, respectively. hHFDCs were passaged three times and subsequently characterized. hHFDCs and MEFs were mitotically inactivated with mitomycin C for 3 h prior to co-culture with H9-hESCs. hESCs were initially established on a mouse feeder layer, subsequently transferred onto a human feeder layer and split every 5 days. Cell morphology, expression of specific 'undifferentiation' markers and growth factors, and the differentiation capacity of hESCs grown on the hHFDC feeder layer were analyzed. hHFDCs are adherent to plastic, possess the classic mesenchymal stem cell phenotype [they express cluster of differentiation (CD)90, CD73 and CD105] and are able to differentiate into adipocytes, chondroblasts and osteocytes, indicating that these cells are multipotent. Population-doubling time analysis revealed that hHFDCs rapidly proliferate over 34.5 h. As a feeder layer, hHFDC behaved similarly to MEF in maintaining the morphology of hESCs. The results of alkaline phosphatase activity, reverse transcription-quantitative polymerase chain reaction analysis of the expression of pluripotency transcription factors [octamer-binding transcription factor 4 (Oct4), Nanog and sex determining region Y-box 2], and immunofluorescence assays of markers (stage-specific embryonic antigen-4 and Oct4) in hESCs co-cultured over hHFDC, indicated that the undifferentiated state of hESCs was preserved. No change in the level of growth factor transcripts (bone morphogenetic protein 4, fibroblast growth factor-2, vascular endothelial growth factor, Pigment epithelium-derived factor and transforming growth factor-β1) was detected for either feeder layer prior to or following inactivation. Similar phenotypes of embryoid body formation, size and morphology were observed in the hHFDC and MEF feeders. In conclusion, hHFDC maintained hESCs in an undifferentiated state comparable to MEF in standard conditions, which may be an important finding regarding the establishment of stem cell-based translational applications.

Authors+Show Affiliations

Instituto de Biofísica Carlos Chagas Filho, Universidade Federal do Rio de Janeiro, Rio de Janeiro, RJ 21941-902, Brazil.Instituto de Biofísica Carlos Chagas Filho, Universidade Federal do Rio de Janeiro, Rio de Janeiro, RJ 21941-902, Brazil.Instituto de Biofísica Carlos Chagas Filho, Universidade Federal do Rio de Janeiro, Rio de Janeiro, RJ 21941-902, Brazil.Instituto de Biofísica Carlos Chagas Filho, Universidade Federal do Rio de Janeiro, Rio de Janeiro, RJ 21941-902, Brazil.Clínica Luiz Pimentel, Niterói, Rio de Janeiro, RJ 24210-480, Brazil.Instituto de Biofísica Carlos Chagas Filho, Universidade Federal do Rio de Janeiro, Rio de Janeiro, RJ 21941-902, Brazil.Instituto de Biofísica Carlos Chagas Filho, Universidade Federal do Rio de Janeiro, Rio de Janeiro, RJ 21941-902, Brazil.Instituto de Biofísica Carlos Chagas Filho, Universidade Federal do Rio de Janeiro, Rio de Janeiro, RJ 21941-902, Brazil.

Pub Type(s)

Journal Article

Language

eng

PubMed ID

28565767

Citation

Coelho de Oliveira, Vanessa Carvalho, et al. "Hair Follicle-derived Mesenchymal Cells Support Undifferentiated Growth of Embryonic Stem Cells." Experimental and Therapeutic Medicine, vol. 13, no. 5, 2017, pp. 1779-1788.
Coelho de Oliveira VC, Silva Dos Santos D, Vairo L, et al. Hair follicle-derived mesenchymal cells support undifferentiated growth of embryonic stem cells. Exp Ther Med. 2017;13(5):1779-1788.
Coelho de Oliveira, V. C., Silva Dos Santos, D., Vairo, L., Kasai Brunswick, T. H., Pimentel, L. A. S., Carvalho, A. B., Campos de Carvalho, A. C., & Goldenberg, R. C. D. S. (2017). Hair follicle-derived mesenchymal cells support undifferentiated growth of embryonic stem cells. Experimental and Therapeutic Medicine, 13(5), 1779-1788. https://doi.org/10.3892/etm.2017.4195
Coelho de Oliveira VC, et al. Hair Follicle-derived Mesenchymal Cells Support Undifferentiated Growth of Embryonic Stem Cells. Exp Ther Med. 2017;13(5):1779-1788. PubMed PMID: 28565767.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Hair follicle-derived mesenchymal cells support undifferentiated growth of embryonic stem cells. AU - Coelho de Oliveira,Vanessa Carvalho, AU - Silva Dos Santos,Danúbia, AU - Vairo,Leandro, AU - Kasai Brunswick,Tais Hanae, AU - Pimentel,Luiz Alberto Soares, AU - Carvalho,Adriana Bastos, AU - Campos de Carvalho,Antonio Carlos, AU - Goldenberg,Regina Coeli Dos Santos, Y1 - 2017/03/08/ PY - 2016/10/14/received PY - 2017/01/13/accepted PY - 2017/6/2/entrez PY - 2017/6/2/pubmed PY - 2017/6/2/medline KW - feeder layer KW - human embryonic stem cell KW - human hair follicle KW - mesenchymal stem cell KW - pluripotency SP - 1779 EP - 1788 JF - Experimental and therapeutic medicine JO - Exp Ther Med VL - 13 IS - 5 N2 - The aim of the present study was to investigate whether feeder layers composed of human hair follicle-derived mesenchymal stem cells (hHFDCs) are able to support human embryonic stem cells (hESCs). hHFDCs and mouse embryonic fibroblasts (MEFs) were isolated and cultured in Dulbecco's modified Eagle's medium (DMEM)/F-12 and low-glucose DMEM, respectively. hHFDCs were passaged three times and subsequently characterized. hHFDCs and MEFs were mitotically inactivated with mitomycin C for 3 h prior to co-culture with H9-hESCs. hESCs were initially established on a mouse feeder layer, subsequently transferred onto a human feeder layer and split every 5 days. Cell morphology, expression of specific 'undifferentiation' markers and growth factors, and the differentiation capacity of hESCs grown on the hHFDC feeder layer were analyzed. hHFDCs are adherent to plastic, possess the classic mesenchymal stem cell phenotype [they express cluster of differentiation (CD)90, CD73 and CD105] and are able to differentiate into adipocytes, chondroblasts and osteocytes, indicating that these cells are multipotent. Population-doubling time analysis revealed that hHFDCs rapidly proliferate over 34.5 h. As a feeder layer, hHFDC behaved similarly to MEF in maintaining the morphology of hESCs. The results of alkaline phosphatase activity, reverse transcription-quantitative polymerase chain reaction analysis of the expression of pluripotency transcription factors [octamer-binding transcription factor 4 (Oct4), Nanog and sex determining region Y-box 2], and immunofluorescence assays of markers (stage-specific embryonic antigen-4 and Oct4) in hESCs co-cultured over hHFDC, indicated that the undifferentiated state of hESCs was preserved. No change in the level of growth factor transcripts (bone morphogenetic protein 4, fibroblast growth factor-2, vascular endothelial growth factor, Pigment epithelium-derived factor and transforming growth factor-β1) was detected for either feeder layer prior to or following inactivation. Similar phenotypes of embryoid body formation, size and morphology were observed in the hHFDC and MEF feeders. In conclusion, hHFDC maintained hESCs in an undifferentiated state comparable to MEF in standard conditions, which may be an important finding regarding the establishment of stem cell-based translational applications. SN - 1792-0981 UR - https://www.unboundmedicine.com/medline/citation/28565767/Hair_follicle_derived_mesenchymal_cells_support_undifferentiated_growth_of_embryonic_stem_cells_ L2 - https://www.ingentaconnect.com/openurl?genre=article&issn=1792-0981&volume=13&issue=5&spage=1779&aulast=Coelho de Oliveira DB - PRIME DP - Unbound Medicine ER -