Isolation and Cryopreservation of Placental Cells: Search for Optimal Biotechniques in Experimental and Regenerative Medicine
Keywords:placenta, cell isolation, explants, cell culture, cryopreservation, mice, rats
High efficacy of placental cells application necessitates their investigation. Preclinical studies require an improvement of the methods for obtaining, standardizing and storage of placental cells of experimental animals. Cells were isolated from rats and mice placentas by means of different enzymatic methods and the one of explants. Cells were cryopreserved with DMSO in DMEM using two-stage freezing. The number, morphological, cultural, metabolic features of cells were studied after isolation and storage. The maximum number of viable cells from the placentas of mice and rats was found to be obtained using the explant method or trypsin with ETDA. Cell cultures from mice and rats placentas after the third passage had stable morphofunctional characteristics. The viability of warmed rat placental cells according to dye exclusion was (92.3 ± 1.6)%, according to the adhesive test this was (81.3 ± 5.8)%. For mice placental cells, these values were (86.7 ± 3.7)% and (79.2 ± 8.1)%, correspondingly. The research results enabled the determining of effective biotechniques for obtaining the cryopreserved placental cells of rats and mice to perform preclinical studies of their biological effect in models of allo- and autotransplantations.
Probl Cryobiol Cryomed 2021; 31(1): 082–088
Araujo AB, Furlan JМ, Salton GD, et al. Isolation of human mesenchymal stem cells from amnion, chorion, placental decidua and umbilical cord: comparison of four enzymatic protocols. Biotechnol Lett. 2018;40(6):989-98. CrossRef
Beeravolu N, McKee C, Alamri A, et al. Isolation and characterization of mesenchymal stromal cells from human umbilical cord and fetal placenta. J Vis Exp [Internet]. 2017 Apr 3 [cited 2019 Dec 10];(122):55224. Available from: https://www.jove.com/pdf/55224/jove-protocol-55224-isolation characterization-mesenchymal-stromal-cells-from-human CrossRef
Choi YS, Park YB, Ha CW, et al. Different characteristics of mesenchymal stem cells isolated from different layers of full term placenta. PLoS ONE [Internet]. 2017 Feb 22 [cited 2019 Dec 10];12(2):e0172642. Available from: https://journals.plos.org/plosone/article?id=10.1371/journal.pone.0172642 CrossRef
Huang Q, Yang Y, Luo C, et al. An efficient protocol to generate placental chorionic plate-derived mesenchymal stem cells with superior proliferative and immunomodulatory properties. Stem Cell Res Ther. 2019;10:301-16. CrossRef
James JL, Hurley DG, Gamage TKJB, et al. Isolation and characterization of a novel trophoblast side-population from first trimester placenta. Reproduction. 2015;150(5):449-62. CrossRef
Kozub MM, Prokopyuk VYu, Skibina KP, et al. Comparison of various of tissue and cell therapy approaches when restoring ovarian, hepatic and kidney's function after chemotherapy induced ovarian failure. Exp Oncol [Internet]. 2017 Sep [cited 2019 Dec 10];39(3):181-5. Available from: https://exp-oncology.com.ua/article/10137 CrossRef
Lobo SE, Leonel LC, Miranda CM, et al. The placenta as an organ and a source of stem cells and extracellular matrix: a review. Cells Tissues Organs. 2016; 201(4):239-52. CrossRef
Miki T, Marongiu F, Ellis E, Strom SC. Isolation of amniotic epithelial stem cells. Curr Protoc Stem Cell Biol [Internet]. 2010 Jan 15 [cited 2019 Dec 10]; Chapter 1: Unit 1E.3. Available from: https://currentprotocols.onlinelibrary.wiley.com/doi/10.1002/9780470151808.sc01e03s3 CrossRef
Naji A, Eitoku M, Favier B, et al. Biological functions of mesenchymal stem cells and clinical implications. Cell Mol Life Sci. 2019;76(17):3323-48. CrossRef
Parolini O, Alviano F, Bagnara GP, et al. Concise review: isolation and characterization of cells from human term placenta: outcome of the first international workshop on placenta derived stem cells. Stem Cells. 2008;6(2):300-11.CrossRef
Pogozhykh D, Pogozhykh O, Prokopyuk V, et al. Influence of temperature fluctuations during cryopreservation on vital parameters, differentiation potential, and transgene expression of placental multipotent stromal cells. Stem Cell Res Ther. [Internet]. 2017 Mar 11 [cited 2019 Dec 10];8(1):66. Available from: https://stemcellres.biomedcentral.com/articles/10.1186/s13287-017-0512-7 CrossRef
Portmann-Lanz CB, Schoeberlein A, Huber A, et al. Placental mesenchymal stem cells as potential autologous graft for pre- and perinatal neuroregeneration. Am J Obstet Gynecol. 2006;194(3):664-73. CrossRef
Prokopyuk OS, Prokopyuk VYu, Pasieshvili NM, et al. Implantation of cryopreserved human placental fragments restores prooxidant-antioxidant balance in experimental animals of late ontogeny. Probl Cryobiol Cryomed. 2017;27(1):61-70. CrossRef
Prokopiuk VYu. Infl uence of media conditioned by cryopreserved and fresh placental explants and cells on murine uterine and ovarian organotypic cultures. Probl Cryobiol Cryomed. 2018;28(2):139-50. CrossRef
Prokopyuk V.Yu., Karpenko V.G., Shevchenko M.V. et al. Experience in clinical application of cryopreserved placental derivatives: cells, tissue, membranes, extract, and cord blood serum. Innov Biosyst Bioeng. 2020; 4 (3): 160-8. CrossRef
Silini AR, Cancelli S, Signoroni PB, et al. The dichotomy of placenta-derived cells in cancer growth. Placenta. 2017;59:154-62. CrossRef
Silini AR, Masserdotti A, Papait A, Parolini O. Shaping the future of perinatal cells: lessons from the past and interpretations of the present. Front Bioeng Biotechnol. [Internet]. 2019 Apr 10 [cited 2019 Dec 10];7:75. Available from: https://www.frontiersin.org/articles/10.3389/fbioe.2019.00075/full CrossRef
Soncini M, Vertua E, Gibelli L, et al. Isolation and characterization of mesenchymal cells from human fetal membranes. J Tissue Eng Regen Med. 2007;1(4):296-305. CrossRef
Svitina H, Kyryk V, Skrypkina I, et al. Placenta-derived multipotent cells have no effect on the size and number of DMH induced colon tumors in rats. Exp Ther Med. 2017;14(3):2135-47. CrossRef
Zhang X, Mitsuru A, Igura K, et al. Mesenchymal progenitor cells derived from chorionic villi of human placenta for cartilage tissue engineering. Biochem Biophys Res Commun. 2006;340(3):944-52. CrossRef
How to Cite
This work is licensed under a Creative Commons Attribution 4.0 International License.
Authors who publish with this journal agree to the following terms:
- Authors retain copyright and grant the journal right of first publication with the work simultaneously licensed under a Creative Commons Attribution License that allows others to share the work with an acknowledgement of the work's authorship and initial publication in this journal.
- Authors are able to enter into separate, additional contractual arrangements for the non-exclusive distribution of the journal's published version of the work (e.g., post it to an institutional repository or publish it in a book), with an acknowledgement of its initial publication in this journal.
- Authors are permitted and encouraged to post their work online (e.g., in institutional repositories or on their website) prior to and during the submission process, as it can lead to productive exchanges, as well as earlier and greater citation of published work (See The Effect of Open Access).