Araştırma Makalesi
Explant culture and multilineage differentiation of amniotic membrane derived stem cells
Yıl 2022, Cilt: 69 Sayı: 2, 191 - 199, 25.03.2022Öz
Amniotic membrane derived stem cells (AMSCs) are reported to have a comparatively higher potency than multipotent stem cells. These cells are shown to have low immunogenicity and no teratogenicity. Among various conventional methods of isolation using enzymes, explant culture method is believed to be an easy and cost-effective way to harvest stem cells. The purpose of this study was to isolate AMSCs from amniotic membrane of rats and to characterize them for multilineage differentiation, including generation of neurospheres to use them later in in-vivo experiments. Amniotic membranes were collected from Wistar rats on 17th day of pregnancy. After processing of the tissues, AMSCs were isolated by the explant culture method and continued to grow until 10th passage. The doubling time was estimated and the cells were analyzed for growth curve parameters at passages 5 and 9. The osteogenic and adipogenic differentiation studies were carried out from the same cells after 3rd passage. Neurospheres generation from AMSCs was performed using neurogenic induction media. The cells were further assessed for their mesenchymal, haemopoietic, and neurogenic marker expressions by immunofluorescence staining and PCR analysis The study suggests that AMSCs isolated through explant culture are reliable stem cells which could generate neurospheres under proper induction conditions and could be a potential candidate to be used on in-vivo neural degeneration models.
Anahtar Kelimeler
Amniotic membrane, differentiation, explant culture, neurosphere
Kaynakça
- Altunbaş K, Yaprakçı MV, Çelik S (2016): Isolation and characterization of olfactory stem cells from canine olfactory mucosa. Kafkas Univ Vet Fak Derg, 22, 237-243.
- Alviano F, Fossati V, Marchionni, et al (2007): Term amniotic membrane is a high throughput source for multipotent mesenchymal stem cells with the ability to differentiate into endothelial cells in vitro. BMC Dev Biol, 7, 11.
- Amendola D, Nardella M, Guglielmi L, et al (2014): Human placenta-derived neurospheres are susceptible to transformation after extensive in vitro expansion. Stem Cell Res Ther, 5, 55.
- Asl KD, Shafaei H, Rad JS, et al (2017): Comparison of characteristics of human amniotic membrane and human adipose tissue derived mesenchymal stem cells. World J Plast Surg, 6, 33.
- Azari H, Louis SA, Sharififar S, et al (2011): Neural-colony forming cell assay: an assay to discriminate bona fide neural stem cells from neural progenitor cells. J Vis Exp, 49, e2639.
- Azedi F, Kazemnejad S, Zarnani AH, et al (2017): Comparative capability of menstrual blood versus bone marrow derived stem cells in neural differentiation. Mol Biol Rep, 44, 169-182.
- Bez A, Corsini E, Curti D, et al (2003): Neurosphere and neurosphere-forming cells: morphological and ultrastructural characterization. Brain Research, 993, 18-29.
- Cardoso MT, Pinheiro AO, Vidane AS, et al (2017): Characterization of teratogenic potential and gene expression in canine and feline amniotic membrane‐derived stem cells. Reprod Domes Anim, 52, 58-64.
- Chung CS, Fujita N, Kawahara N, et al (2013): A comparison of neurosphere differentiation potential of canine bone marrow-derived mesenchymal stem cells and adipose-derived mesenchymal stem cells. J Vet Med Sci, 75, 879-886.
- Corradetti B, Meucci A, Bizzaro D, et al (2013): Mesenchymal stem cells from amnion and amniotic fluid in the bovine. Reproduction, 145, 391-400.
- Cremonesi F, Corradetti B, Consiglio AL (2011): Fetal adnexa derived stem cells from domestic animal: progress and perspectives. Theriogenology, 75, 1400-1415.
- Díaz-Prado S, Muiños-López E, Hermida-Gómez T, et al (2010): Isolation and characterization of mesenchymal stem cells from human amniotic membrane. Tissue Eng Part C: Methods, 17, 49-59.
- Fukuchi Y, Nakajima H, Sugiyama D, et al (2004): Human placenta‐derived cells have mesenchymal stem/progenitor cell potential. Stem Cells, 22, 649-658.
- Girard SD, Devéze A, Nivet E, et al (2011): Isolating nasal olfactory stem cells from rodents or humans. J Vis Exp, 54, e2762.
- Jing W, Xiao J, Xiong Z, et al (2011): Explant culture: an efficient method to isolate adipose‐derived stromal cells for tissue engineering. Artif Organs, 35, 105-112.
- Kalendar R, Lee D, Schulman AH (2009): FastPCR software for PCR primer and probe design and repeat search. G3, 3, 1-14.
- Kim J, Lee Y, Kim H, et al (2007): Human amniotic fluid‐derived stem cells have characteristics of multipotent stem cells. Cell Prolif, 40, 75-90.
- Lange‐Consiglio A, Corradetti B, Bizzaro D, et al (2012): Characterization and potential applications of progenitor‐like cells isolated from horse amniotic membrane. J Tissue Eng Regen Med, 6, 622-635.
- Lange‐Consiglio A, Corradetti B, Meucci A, et al (2013): Characteristics of equine mesenchymal stem cells derived from amnion and bone marrow: in vitro proliferative and multilineage potential assessment. Equine Vet J, 45, 737-744.
- Lange‐Consiglio A, Corradetti B, Bertani S, et al (2015): Peculiarity of porcine amniotic membrane and its derived cells: a contribution to the study of cell therapy from a large animal model. Cell Reprogram, 17, 472-483.
- Lapchak PA (2015): A cost-effective rabbit embolic stroke bioassay: insight into the development of acute ischemic stroke therapy. Transl Stroke Res, 6, 99-103.
- Li H, Yu S, Hao F, et al (2018): Insulin‐like growth factor binding protein 4 inhibits proliferation of bone marrow mesenchymal stem cells and enhances growth of neurospheres derived from the stem cells. Cell Biochem Funct, 36, 331-341.
- Lobo MV, Alonso FJM, Redondo C, et al (2003): Cellular characterization of epidermal growth factor-expanded free-floating neurospheres. J Histochem Cytochem, 51, 89-103.
- Magatti M, Vertua E, Cargnoni A, et al (2018): The immunomodulatory properties of amniotic cells: the two sides of the coin. Cell Transplant, 27, 31-44.
- Marcus AJ, Coyne TM, Rauch J, et al (2008): Isolation, characterization, and differentiation of stem cells derived from the rat amniotic membrane. Differentiation, 76, 130-144.
- Mihu CM, Rus-Ciuca D, Soritau O, et al (2009): Isolation and characterization of mesenchymal stem cells from the amniotic membrane. Rom J Morphol Embryol, 50, 73-77.
- Nawaz S, Özden Akkaya Ö, Dikmen T, et al (2020): Molecular characterization of bovine amniotic fluid derived stem cells with an underlying focus on their comparative neuronal potential at different passages. Ann Anat, 228, 151452.
- Özden-Akkaya Ö, Dikmen T, Nawaz S (2019): Investigation of Sox2, ß-III tubulinand nestin expressions in neuropsheres differentiated from bovine adipose derived mesenchymal stem cells by immunofluorescence staining. Kocatepe Vet J, 12, 336-342.
- Pirjali T, Azarpira N, Ayatollahi M, et al (2013): Isolation and characterization of human mesenchymal stem cells derived from human umbilical cord Wharton’s jelly and amniotic membrane. Int J Organ Transplant Med, 4, 111-116.
- Portmann-Lanz CB, Schoeberlein A, Huber A, et al (2006): Placental mesenchymal stem cells as potential autologous graft for pre-and perinatal neuroregeneration. Am J Obstet Gynecol, 194, 664-673.
- Reynolds BA, Weiss S (1992): Generation of neurons and astrocytes from isolated cells of the adult mammalian central nervous system. Science, 255, 1707-1710.
- Rezaei F, Tiraihi T, Abdanipour A, et al (2018): Immunocytochemical analysis of valproic acid induced histone H3 and H4 acetylation during differentiation of rat adipose derived stem cells into neuron-like cells. Biotech & Histochem, 93, 589-600.
- Roth V (2006): Doubling Time Computing, Available from: http://www.doubling-time.com/compute.php. (Accessed Nov 11, 2020).
- Rutigliano L, Corradetti B, Valentini L, et al (2013): Molecular characterization and in vitro differentiation of feline progenitor-like amniotic epithelial cells. Stem Cell Res Ther, 4, 133.
- Salehinejad P, Alitheen NB, Ali AM, et al (2012): Comparison of different methods for the isolation of mesenchymal stem cells from human umbilical cord Wharton’s jelly. In Vitro Cell Dev Biol Anim, 48, 75-83.
- Sarugaser R, Lickorish D, Baksh D, et al (2005): Human umbilical cord perivascular (HUCPV) cells: a source of mesenchymal progenitors. Stem Cells, 23, 220-229.
- Seo MS, Park SB, Kim HS, et al (2013): Isolation and characterization of equine amniotic membrane-derived mesenchymal stem cells. J Vet Sci, 14, 151-159.
- Sun J, Wei ZZ, Gu X, et al (2015): Intranasal delivery of hypoxia-preconditioned bone marrow-derived mesenchymal stem cells enhanced regenerative effects after intracerebral hemorrhagic stroke in mice. Exp Neurol, 272, 78-87.
- Suslov ON, Kukekov VG, Ignatova TN, et al (2002): Neural stem cell heterogeneity demonstrated by molecular phenotyping of clonal neurospheres. PNAS, 99, 14506-14511.
- Tang Y, Zhang C, Wang J, et al (2015): MRI/SPECT/fluorescent tri‐modal probe for evaluating the homing and therapeutic efficacy of transplanted mesenchymal stem cells in a rat ischemic stroke model. Adv Funct Mater, 25, 1024-1034.
- Woodbury D, Marcus AJ (2015): Obtaining multipotent amnion-derived stem cell (ADSC) from amniotic membrane tissue without enzymatic digestion. Google Patents.
- Yan ZJ, Zhang P, Hu YQ, et al (2013): Neural stem-like cells derived from human amnion tissue are effective in treating traumatic brain injury in rat. Neurochem Res, 38, 1022-1033.
- Yoon JH, Roh EY, Shin S, et al (2013): Comparison of explant-derived and enzymatic digestion-derived MSCs and the growth factors from Wharton’s jelly. Bio Med Res Int, 2013, 428726.
- Zhou HL, Zhang XJ, Zhang MY, et al (2016): Transplantation of human amniotic mesenchymal stem cells promotes functional recovery in a rat model of traumatic spinal cord injury. Neurochem Res, 41, 2708-2718.
Yıl 2022, Cilt: 69 Sayı: 2, 191 - 199, 25.03.2022
Öz
Kaynakça
- Altunbaş K, Yaprakçı MV, Çelik S (2016): Isolation and characterization of olfactory stem cells from canine olfactory mucosa. Kafkas Univ Vet Fak Derg, 22, 237-243.
- Alviano F, Fossati V, Marchionni, et al (2007): Term amniotic membrane is a high throughput source for multipotent mesenchymal stem cells with the ability to differentiate into endothelial cells in vitro. BMC Dev Biol, 7, 11.
- Amendola D, Nardella M, Guglielmi L, et al (2014): Human placenta-derived neurospheres are susceptible to transformation after extensive in vitro expansion. Stem Cell Res Ther, 5, 55.
- Asl KD, Shafaei H, Rad JS, et al (2017): Comparison of characteristics of human amniotic membrane and human adipose tissue derived mesenchymal stem cells. World J Plast Surg, 6, 33.
- Azari H, Louis SA, Sharififar S, et al (2011): Neural-colony forming cell assay: an assay to discriminate bona fide neural stem cells from neural progenitor cells. J Vis Exp, 49, e2639.
- Azedi F, Kazemnejad S, Zarnani AH, et al (2017): Comparative capability of menstrual blood versus bone marrow derived stem cells in neural differentiation. Mol Biol Rep, 44, 169-182.
- Bez A, Corsini E, Curti D, et al (2003): Neurosphere and neurosphere-forming cells: morphological and ultrastructural characterization. Brain Research, 993, 18-29.
- Cardoso MT, Pinheiro AO, Vidane AS, et al (2017): Characterization of teratogenic potential and gene expression in canine and feline amniotic membrane‐derived stem cells. Reprod Domes Anim, 52, 58-64.
- Chung CS, Fujita N, Kawahara N, et al (2013): A comparison of neurosphere differentiation potential of canine bone marrow-derived mesenchymal stem cells and adipose-derived mesenchymal stem cells. J Vet Med Sci, 75, 879-886.
- Corradetti B, Meucci A, Bizzaro D, et al (2013): Mesenchymal stem cells from amnion and amniotic fluid in the bovine. Reproduction, 145, 391-400.
- Cremonesi F, Corradetti B, Consiglio AL (2011): Fetal adnexa derived stem cells from domestic animal: progress and perspectives. Theriogenology, 75, 1400-1415.
- Díaz-Prado S, Muiños-López E, Hermida-Gómez T, et al (2010): Isolation and characterization of mesenchymal stem cells from human amniotic membrane. Tissue Eng Part C: Methods, 17, 49-59.
- Fukuchi Y, Nakajima H, Sugiyama D, et al (2004): Human placenta‐derived cells have mesenchymal stem/progenitor cell potential. Stem Cells, 22, 649-658.
- Girard SD, Devéze A, Nivet E, et al (2011): Isolating nasal olfactory stem cells from rodents or humans. J Vis Exp, 54, e2762.
- Jing W, Xiao J, Xiong Z, et al (2011): Explant culture: an efficient method to isolate adipose‐derived stromal cells for tissue engineering. Artif Organs, 35, 105-112.
- Kalendar R, Lee D, Schulman AH (2009): FastPCR software for PCR primer and probe design and repeat search. G3, 3, 1-14.
- Kim J, Lee Y, Kim H, et al (2007): Human amniotic fluid‐derived stem cells have characteristics of multipotent stem cells. Cell Prolif, 40, 75-90.
- Lange‐Consiglio A, Corradetti B, Bizzaro D, et al (2012): Characterization and potential applications of progenitor‐like cells isolated from horse amniotic membrane. J Tissue Eng Regen Med, 6, 622-635.
- Lange‐Consiglio A, Corradetti B, Meucci A, et al (2013): Characteristics of equine mesenchymal stem cells derived from amnion and bone marrow: in vitro proliferative and multilineage potential assessment. Equine Vet J, 45, 737-744.
- Lange‐Consiglio A, Corradetti B, Bertani S, et al (2015): Peculiarity of porcine amniotic membrane and its derived cells: a contribution to the study of cell therapy from a large animal model. Cell Reprogram, 17, 472-483.
- Lapchak PA (2015): A cost-effective rabbit embolic stroke bioassay: insight into the development of acute ischemic stroke therapy. Transl Stroke Res, 6, 99-103.
- Li H, Yu S, Hao F, et al (2018): Insulin‐like growth factor binding protein 4 inhibits proliferation of bone marrow mesenchymal stem cells and enhances growth of neurospheres derived from the stem cells. Cell Biochem Funct, 36, 331-341.
- Lobo MV, Alonso FJM, Redondo C, et al (2003): Cellular characterization of epidermal growth factor-expanded free-floating neurospheres. J Histochem Cytochem, 51, 89-103.
- Magatti M, Vertua E, Cargnoni A, et al (2018): The immunomodulatory properties of amniotic cells: the two sides of the coin. Cell Transplant, 27, 31-44.
- Marcus AJ, Coyne TM, Rauch J, et al (2008): Isolation, characterization, and differentiation of stem cells derived from the rat amniotic membrane. Differentiation, 76, 130-144.
- Mihu CM, Rus-Ciuca D, Soritau O, et al (2009): Isolation and characterization of mesenchymal stem cells from the amniotic membrane. Rom J Morphol Embryol, 50, 73-77.
- Nawaz S, Özden Akkaya Ö, Dikmen T, et al (2020): Molecular characterization of bovine amniotic fluid derived stem cells with an underlying focus on their comparative neuronal potential at different passages. Ann Anat, 228, 151452.
- Özden-Akkaya Ö, Dikmen T, Nawaz S (2019): Investigation of Sox2, ß-III tubulinand nestin expressions in neuropsheres differentiated from bovine adipose derived mesenchymal stem cells by immunofluorescence staining. Kocatepe Vet J, 12, 336-342.
- Pirjali T, Azarpira N, Ayatollahi M, et al (2013): Isolation and characterization of human mesenchymal stem cells derived from human umbilical cord Wharton’s jelly and amniotic membrane. Int J Organ Transplant Med, 4, 111-116.
- Portmann-Lanz CB, Schoeberlein A, Huber A, et al (2006): Placental mesenchymal stem cells as potential autologous graft for pre-and perinatal neuroregeneration. Am J Obstet Gynecol, 194, 664-673.
- Reynolds BA, Weiss S (1992): Generation of neurons and astrocytes from isolated cells of the adult mammalian central nervous system. Science, 255, 1707-1710.
- Rezaei F, Tiraihi T, Abdanipour A, et al (2018): Immunocytochemical analysis of valproic acid induced histone H3 and H4 acetylation during differentiation of rat adipose derived stem cells into neuron-like cells. Biotech & Histochem, 93, 589-600.
- Roth V (2006): Doubling Time Computing, Available from: http://www.doubling-time.com/compute.php. (Accessed Nov 11, 2020).
- Rutigliano L, Corradetti B, Valentini L, et al (2013): Molecular characterization and in vitro differentiation of feline progenitor-like amniotic epithelial cells. Stem Cell Res Ther, 4, 133.
- Salehinejad P, Alitheen NB, Ali AM, et al (2012): Comparison of different methods for the isolation of mesenchymal stem cells from human umbilical cord Wharton’s jelly. In Vitro Cell Dev Biol Anim, 48, 75-83.
- Sarugaser R, Lickorish D, Baksh D, et al (2005): Human umbilical cord perivascular (HUCPV) cells: a source of mesenchymal progenitors. Stem Cells, 23, 220-229.
- Seo MS, Park SB, Kim HS, et al (2013): Isolation and characterization of equine amniotic membrane-derived mesenchymal stem cells. J Vet Sci, 14, 151-159.
- Sun J, Wei ZZ, Gu X, et al (2015): Intranasal delivery of hypoxia-preconditioned bone marrow-derived mesenchymal stem cells enhanced regenerative effects after intracerebral hemorrhagic stroke in mice. Exp Neurol, 272, 78-87.
- Suslov ON, Kukekov VG, Ignatova TN, et al (2002): Neural stem cell heterogeneity demonstrated by molecular phenotyping of clonal neurospheres. PNAS, 99, 14506-14511.
- Tang Y, Zhang C, Wang J, et al (2015): MRI/SPECT/fluorescent tri‐modal probe for evaluating the homing and therapeutic efficacy of transplanted mesenchymal stem cells in a rat ischemic stroke model. Adv Funct Mater, 25, 1024-1034.
- Woodbury D, Marcus AJ (2015): Obtaining multipotent amnion-derived stem cell (ADSC) from amniotic membrane tissue without enzymatic digestion. Google Patents.
- Yan ZJ, Zhang P, Hu YQ, et al (2013): Neural stem-like cells derived from human amnion tissue are effective in treating traumatic brain injury in rat. Neurochem Res, 38, 1022-1033.
- Yoon JH, Roh EY, Shin S, et al (2013): Comparison of explant-derived and enzymatic digestion-derived MSCs and the growth factors from Wharton’s jelly. Bio Med Res Int, 2013, 428726.
- Zhou HL, Zhang XJ, Zhang MY, et al (2016): Transplantation of human amniotic mesenchymal stem cells promotes functional recovery in a rat model of traumatic spinal cord injury. Neurochem Res, 41, 2708-2718.
Ayrıntılar
| Birincil Dil | İngilizce |
|---|---|
| Konular | Veteriner Hekimlik |
| Bölüm | Araştırma Makalesi |
| Yazarlar |
|
| Destekleyen Kurum | This work was supported by Afyon Kocatepe University, Scientific Research Projects Coordination Unit (16.KARİYER.23), Afyonkarahisar, Turkey. |
| Proje Numarası | 16.KARİYER.23 |
| Teşekkür | We would like to acknowledge Mr. Tayfun Dikmen for his contributions in editing this article. A part of this study was presented in 3rd International Vetistanbul Group Congress, May 17-20 2016, Sarajevo, Bosnia and Herzegovina. |
| Yayımlanma Tarihi | 25 Mart 2022 |
| Yayınlandığı Sayı | Yıl 2022Cilt: 69 Sayı: 2 |
Kaynak Göster
| Bibtex | @araştırma makalesi { auvfd830460, journal = {Ankara Üniversitesi Veteriner Fakültesi Dergisi}, issn = {1300-0861}, eissn = {1308-2817}, address = {}, publisher = {Ankara Üniversitesi}, year = {2022}, volume = {69}, number = {2}, pages = {191 - 199}, doi = {10.33988/auvfd.830460}, title = {Explant culture and multilineage differentiation of amniotic membrane derived stem cells}, key = {cite}, author = {Özden Akkaya, Özlem and Kıbrıa, Asm Golam and Yağcı, Artay and Erdoğan, Metin and Altunbaş, Korhan} } |
| APA | Özden Akkaya, Ö. , Kıbrıa, A. G. , Yağcı, A. , Erdoğan, M. & Altunbaş, K. (2022). Explant culture and multilineage differentiation of amniotic membrane derived stem cells . Ankara Üniversitesi Veteriner Fakültesi Dergisi , 69 (2) , 191-199 . DOI: 10.33988/auvfd.830460 |
| MLA | Özden Akkaya, Ö. , Kıbrıa, A. G. , Yağcı, A. , Erdoğan, M. , Altunbaş, K. "Explant culture and multilineage differentiation of amniotic membrane derived stem cells" . Ankara Üniversitesi Veteriner Fakültesi Dergisi 69 (2022 ): 191-199 <http://vetjournal.ankara.edu.tr/tr/pub/issue/69048/830460> |
| Chicago | Özden Akkaya, Ö. , Kıbrıa, A. G. , Yağcı, A. , Erdoğan, M. , Altunbaş, K. "Explant culture and multilineage differentiation of amniotic membrane derived stem cells". Ankara Üniversitesi Veteriner Fakültesi Dergisi 69 (2022 ): 191-199 |
| RIS | TY - JOUR T1 - Explant culture and multilineage differentiation of amniotic membrane derived stem cells AU - ÖzlemÖzden Akkaya, Asm GolamKıbrıa, ArtayYağcı, MetinErdoğan, KorhanAltunbaş Y1 - 2022 PY - 2022 N1 - doi: 10.33988/auvfd.830460 DO - 10.33988/auvfd.830460 T2 - Ankara Üniversitesi Veteriner Fakültesi Dergisi JF - Journal JO - JOR SP - 191 EP - 199 VL - 69 IS - 2 SN - 1300-0861-1308-2817 M3 - doi: 10.33988/auvfd.830460 UR - https://doi.org/10.33988/auvfd.830460 Y2 - 2021 ER - |
| EndNote | %0 Ankara Üniversitesi Veteriner Fakültesi Dergisi Explant culture and multilineage differentiation of amniotic membrane derived stem cells %A Özlem Özden Akkaya , Asm Golam Kıbrıa , Artay Yağcı , Metin Erdoğan , Korhan Altunbaş %T Explant culture and multilineage differentiation of amniotic membrane derived stem cells %D 2022 %J Ankara Üniversitesi Veteriner Fakültesi Dergisi %P 1300-0861-1308-2817 %V 69 %N 2 %R doi: 10.33988/auvfd.830460 %U 10.33988/auvfd.830460 |
| ISNAD | Özden Akkaya, Özlem , Kıbrıa, Asm Golam , Yağcı, Artay , Erdoğan, Metin , Altunbaş, Korhan . "Explant culture and multilineage differentiation of amniotic membrane derived stem cells". Ankara Üniversitesi Veteriner Fakültesi Dergisi 69 / 2 (Mart 2022): 191-199 . https://doi.org/10.33988/auvfd.830460 |
| AMA | Özden Akkaya Ö. , Kıbrıa A. G. , Yağcı A. , Erdoğan M. , Altunbaş K. Explant culture and multilineage differentiation of amniotic membrane derived stem cells. Ankara Univ Vet Fak Derg. 2022; 69(2): 191-199. |
| Vancouver | Özden Akkaya Ö. , Kıbrıa A. G. , Yağcı A. , Erdoğan M. , Altunbaş K. Explant culture and multilineage differentiation of amniotic membrane derived stem cells. Ankara Üniversitesi Veteriner Fakültesi Dergisi. 2022; 69(2): 191-199. |
| IEEE | Ö. Özden Akkaya , A. G. Kıbrıa , A. Yağcı , M. Erdoğan ve K. Altunbaş , "Explant culture and multilineage differentiation of amniotic membrane derived stem cells", Ankara Üniversitesi Veteriner Fakültesi Dergisi, c. 69, sayı. 2, ss. 191-199, Mar. 2022, doi:10.33988/auvfd.830460 |
