Isolation and Characterization of Fibroblast from Normal and Thalassemia Foreskin
DOI:
https://doi.org/10.48048/tis.2024.7672Keywords:
Characterization, Fibroblast, Flow cytometry, Isolation, ThalassemiaAbstract
Thalassemia is a genetic blood disorder impacting hemoglobin production, varies in severity, and requires lifelong treatments. The advancement in somatic cell reprogramming through induced pluripotent stem cells (iPSCs) represents a personalized medicine approach that holds promise as a treatment for individuals with thalassemia. One source that could be reprogrammed into iPSCs can be generated from foreskin fibroblast cells. This study aimed to isolate and characterize human fibroblast cells from normal (NFF) and thalassemia (TFF) foreskin surface markers (CD44, CD90, CD105, CD73), and negative lineage (CD45, CD34, CD11b, CD19, HLA-DR). Fibroblast cells were isolated out of normal and thalassemia foreskin using the explant method. Characterization of NFF and TFF isolates was carried out using flow cytometry to disclose the expression of CD90, CD44, CD73, CD105, and negative lineage. Isolation of fibroblast cells from normal and thalassemia foreskin was successfully carried out using the explant method with NFF and TFF characterized as expressing CD90, CD44, CD105, CD73, and negative lineages (CD45, CD11b, CD34, CD19, and HLA-DR). This result could lead to a continuation of reprogramming into iPSCs.
HIGHLIGHTS
- Isolation of fibroblast cells from normal and thalassemia foreskin was successfully carried out and cultured within 21 days
- The surface marker (CD44, CD73, CD90, CD105), and negative lineage (CD45, CD34, CD11b, CD19, HLA-DR) showed similarity to MSCs
- Fibroblast cells from the foreskin of normal and thalassemia patients were detected expressing CD90, CD105, CD73, CD44, and negative lineages (CD34, CD11b, CD45, CD19, HLA-DR)
GRAPHICAL ABSTRACT
Downloads
Metrics
References
MA Meri, AH Al-Hakeem and RS Al-Abeadi. An overview on thalassemia: A review article. Med. Sci. J. Adv. Res. 2022; 3, 26-32.
Z Rustam, A Kamalia, R Hidayat, F Subroto and A Suryansyah. Comparison of fuzzy c-means, fuzzy kernel c-means, and fuzzy kernel robust c-means to classify thalassemia data. Int. J. Adv. Sci. Eng. Inform. Tech. 2019; 9, 1205-10.
E Vichinsky. Non-transfusion-dependent thalassemia and thalassemia intermedia: Epidemiology, complications, and management. Curr. Med. Res. Opin. 2016; 32, 191-204.
K Torcharus and T Pankaew. Health-related quality of life in Thai thalassemic children treated with iron chelation. Southeast Asian J. Trop. Med. Publ. Health 2011; 42, 951-9.
S Sharma, B Seth, P Jawade, M Ingale and MS Setia. Quality of life in children with thalassemia and their caregivers in India. Indian J. Pediatr. 2017; 84, 188-94.
KM Musallam, L Lombard, KD Kistler, M Arregui, KS Gilroy, C Chamberlain, E Zagadailov, K Ruiz and AT Taher. Epidemiology of clinically significant forms of alpha‐and beta‐thalassemia: A global map of evidence and gaps. Am. J. Hematol. 2023; 98, 1436-51.
FR Aszhari, Z Rustam, F Subroto and AS Semendawai. Classification of thalassemia data using random forest algorithm. J. Phys. Conf. Ser. 2020; 1490, 1-7.
AA Shafie, IK Chhabra, JHY Wong, NS Mohammed, HM Ibrahim and H Alias. Health-related quality of life among children with transfusion-dependent thalassemia: A cross-sectional study in Malaysia. Health Qual. Life Outcome. 2020; 18, 1-11.
M Faranoush, P Faranoush, I Heydari, MR Foroughi‐Gilvaee, A Azarkeivan, AP Kia, N Sadighnia, A Elahinia, A Zandi, MR Rezvany, N Hashemi-Madani, A Ziaee, R Nekouian and F Rohani. Complications in patients with transfusion dependent thalassemia: A descriptive cross‐sectional study. Health Sci. Rep. 2023; 6, 1-10.
C Borgna-Pignatti, M Marsella. Iron chelation in thalassemia major. Clin Ther. 2015; 37: 2866-77.
Y Wang, CG Zheng, Y Jiang, J Zhang, J Chen, C Yao, Q Zhao, S Liu, K Chen, J Du, Z Yang and S Gao. Genetic correction of β-thalassemia patient-specific iPS cells and its use in improving hemoglobin production in irradiated SCID mice. Cell Res. 2012; 22, 637-48.
T Oliveira, I Costa, V Marinho, V Carvalho, K Uchôa, C Ayres, S Teixeira and DFP Vasconcelos. Human foreskin fibroblasts: From waste bag to important biomedical applications. J. Clin. Urol. 2018; 11, 385-94.
MV Plikus, X Wang, S Sinha, E Forte, SM Thompson, EL Herzog, RR Driskell, N Rosenthal, J Biernaskie and V Horsley. Fibroblasts: Origins, definitions, and functions in health and disease. Cell 2021; 184, 3852-72.
RA Denu, S Nemcek, DD Bloom, A Goodrich, J Kim, DF Mosher and P Hematti. Fibroblasts and mesenchymal stromal/stem cells are phenotypically indistinguishable. Acta Haematol. 2016; 136, 85-97.
J Zupan. Mesenchymal stem/stromal cells and fibroblasts: Their roles in tissue injury and regeneration, and age-related degeneration. IntechOpen, London, 2021.
L Hananta, L Astuti, AH Sadewa, J Alice and J Hutagalung. The prevalence of CYP2B6 gene polymorphisms in malaria-endemic population of Timor in East Nusa Tenggara Indonesia. Osong Publ. Health Res. Perspect. 2018; 9, 192-6.
W Widowati, RF Gunanegara, R Rizal, WS Widodo, A Amalia, SHB Wibowo, K Handono, M Marlina, INE Lister and L Chiuman. Comparative analysis of Wharton’s Jelly mesenchymal stem cell (WJ-MSCs) isolated using explant and enzymatic methods. J. Phys. Conf. Ser. 2019; 1374, 1-11.
AA Antoninus, W Widowati, L Wijaya, D Agustina, S Puradisastra, SB Sumitro, MA Widodo and I Bachtiar. Human platelet lysate enhances the proliferation of Wharton’s jelly-derived mesenchymal stem cells. Biomarkers Genom. Med. 2015; 7, 87-97.
F Nejaddehbashi, V Bayati, L Mashali, M Hashemitabar, M Abbaspour, E Moghimipour and M Orazizadeh. Isolating human dermal fibroblasts using serial explant culture. Stem Cell Investig. 2019; 6, 1-8.
MG Jagadeeshaprasad, PK Govindappa, AM Nelson and JC Elfar. Isolation, culture, and characterization of primary schwann cells, keratinocytes, and fibroblasts from human foreskin. J. Vis. Exp. 2022; 23, e63776.
W Widowati, R Noverina, W Ayuningtyas, D Kurniawan, HSW Kusuma, S Arumwardana, DS Artie, IA Sholihah, RrAS Handayani, DR Laksmitawati, R Rinendyaputri, R Rilianawati and A Faried. Proliferation, characterization, and differentiation potency of adipose tissue-derived mesenchymal stem cells (AT-MSCs) cultured in fresh frozen and non-fresh frozen plasma. Int. J. Mol. Cell. Med. 2019b; 8, 283-94.
K Takahashi, K Tanabe, M Ohnuki, M Narita, T Ichisaka, K Tomoda and S Yamanaka. Induction of pluripotent stem cells from adult human fibroblasts by defined factors. Cell 2007; 131, 861‐72.
RL Thangapazham, TN Darling and J Meyerle. Alteration of skin properties with autologous dermal fibroblasts. Int. J. Mol. Sci. 2014; 15, 8407-27.
AM Sacco, L Belviso, V Romano, A Carfora, F Schonauer, D Nurzynska, S Montagnani, FD Meglio and C Castaldo. Diversity of dermal fibroblasts as major determinant of variability in cell reprogramming. J. Cell. Mol. Med. 2019; 23, 4256-68.
R Marcoccia, S Nesci, B Merlo, G Ballotta, C Algieri, A Pagliarani and E Iacono. Biological characteristics and metabolic profile of canine mesenchymal stem cells isolated from adipose tissue and umbilical cord matrix. PLoS One 2021; 16, 1-19.
DA Moraes, TT Sibov, LF Pavon, PQ Alvim, RS Bonadio, JR Da Silva, A Pic-Taylor, OA Toledo, LC Marti, RB Azevedo and DM Oliveira. A reduction in CD90 (THY-1) expression results in increased differentiation of mesenchymal stromal cells. Stem Cell Res. Ther. 2016; 7, 1-14.
LACD Carvalho, SLTD Santos, LV Sacramento, VRDA Júnior, FCDA Xavier, JND Santos and ACGH Leitão. Mesenchymal stem cell markers in periodontal tissues and periapical lesions. Acta Histochem. 2020; 122, 1-10.
C Brown, C McKee, S Bakshi, K Walker, E Hakman, S Halassy, D Svinarich, R Dodds, CK Govind and GR Chaudry. Mesenchymal stem cells: Cell therapy and regeneration potential. J. Tissue Eng. Regen Med. 2019; 13, 1738-55.
J Wang, Z Chen, M Sun, H Xu, Y Gao, J Liu, M Li. Characterization and therapeutic applications of mesenchymal stem cells for regenerative medicine. Tissue Cell. 2020; 64, 101330.
TA Krylova, AS Musorina, AM Koltsova, VV Zenin, VL Turilova, TK Yakovleva, GG Poljanskaya, Isolation and comparative characteristics of mesenchymal stem-cell lines derived from foreskin of two donors of similar age. Cell Tissue Biol. 2018; 12, 271-80.
TS Dutt, SM LaVergne, TL Webb, BA Baxter, S Stromberg, K McFann, K Berry, M Tipton, O Alnachoukati, L Zier, G Ebel, J Dunn, M Henao-Tamayo and EP Ryan. Comprehensive immune profiling reveals CD56+ monocytes and CD31+ endothelial cells are increased in severe COVID-19 disease. J. Immunol. 2022; 208, 685-96.
Downloads
Published
How to Cite
Issue
Section
License
Copyright (c) 2024 Walailak University
This work is licensed under a Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International License.
