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Establishment and characterization of a fibroblast cell line from postmortem skin of an adult Chinese muntjac (Muntiacus reevesi)

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Abstract

Isolation and culture of somatic cells from animals especially endangered species have raised great concerns as it is being an effective and convenient way to preserve genetic materials for future studies. As a species native to China, Chinese muntjac (Muntiacus reevesi) is listed as a beneficial species with economic and scientific research values. To our knowledge, however, there have been no published reports on somatic cell preservation of this species to date. To conserve biological resources for sustainability of Chinese muntjacs’ genetic diversity, we established a fibroblast cell line from the postmortem ear skin of an adult male Chinese muntjac. The cultured cells were adherent to the plastic and showed an elongated, thin, and spindle-like shape. Moreover, they were FSP1- and VIM-positive characterizing them to be fibroblastic. No microorganisms (bacteria, fungi, or mycoplasmas) were detected throughout the whole study. Cell viability was high although it declined somehow after passaging. The population doubling time was 21.28 h according to the growth curve. Chromosome analysis revealed that the established fibroblast cell line contained 23 pairs of chromosomes, one pair of which was sex chromosomes (XY). Mitochondrial cytochrome C oxidase I gene of cultured cells shared 98.32% identity with those of Muntiacus reevesi registered in GenBank, which verified the cell line was derived from Muntiacus reevesi. In conclusion, we propagated and characterized fibroblast cells from a Chinese muntjac. We believe that this somatic cell line could facilitate animal cloning and breeding studies and become a useful in vitro model to address genetic questions.

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References

  • Chi J, Fu B, Nie W, Wang J, Graphodatsky AS, Yang F (2005) New insights into the karyotypic relationships of Chinese muntjac (Muntiacus reevesi), forest musk deer (Moschus berezovskii) and gayal (Bos frontalis). Cytogenet Genome Res 108:310–316

    Article  CAS  Google Scholar 

  • Chiang P, Lin CC, Liao S, Hsieh L, Li S, Chao M, Li Y (2004) Genetic analysis of two subspecies of Reeves' Muntjac (Cervidae: Muntiacus reevesi) by karyotyping and satellite DNA analyses. Zool Stud 43:749–758

    CAS  Google Scholar 

  • Daneshvar Amoli A, Mohebali N, Farzaneh P, Shahzadeh Fazeli SA, Nikfarjam L, Ashouri Movasagh S, Moradmand Z, Ganjibakhsh M, Nasimian A, Izadpanah M, Vakhshiteh F, Gohari NS, Masoudi NS, Farghadan M, Mohamadi Moghanjoghi S, Khalili M, Khaledi KJ (2017) Establishment and characterization of Caspian horse fibroblast cell bank in Iran. In Vitro Cell Dev Biol Anim 53:337–343

    Article  CAS  Google Scholar 

  • Folmer O, Black M, Hoeh W, Lutz R, Vrijenhoek R (1994) DNA primers for amplification of mitochondrial cytochrome c oxidase subunit I from diverse metazoan invertebrates. Mol Mar Biol Biotechnol 3:294–299

    CAS  Google Scholar 

  • Frönicke L, Chowdhary BP, Scherthan H (1997) Segmental homology among cattle (Bos taurus), Indian muntjac (Muntiacus muntjak vaginalis), and Chinese muntjac (M. reevesi) karyotypes. Cytogenet Cell Genet 77:223–227

    Article  Google Scholar 

  • Hebert PDN, Ratnasingham S, de Waard JR (2003) Barcoding animal life: cytochrome c oxidase subunit 1 divergences among closely related species. Proc Biol Sci 270(Suppl 1):S96–S99

    CAS  PubMed  PubMed Central  Google Scholar 

  • Johnston FP, Church RB, Lin CC (1982) Chromosome rearrangement between the Indian muntjac and Chinese muntjac is accompanied by a deletion of middle repetitive DNA. Can J Biochem 60:497–506

    Article  CAS  Google Scholar 

  • Li XC, Yue H, Li CY, He XH, Zhao QJ, Ma YH, Guan WJ, Ma JZ (2009) Establishment and characterization of a fibroblast cell line derived from Jining black grey goat for genetic conservation. Small Rumin Res 87:17–26

    Article  Google Scholar 

  • Machado LC, Oliveira VC, Paraventi MD, Cardoso RNR, Martins DS, Ambrósio CE (2016) Maintenance of Brazilian biodiversity by germplasm bank. Pesqui Vet Bras 36:62–66

    Article  Google Scholar 

  • Moro LN, Hiriart MI, Buemo C, Jarazo J, Sestelo A, Veraguas D, Rodriguez-Alvarez L, Salamone DF (2015a) Cheetah interspecific SCNT followed by embryo aggregation improves in vitro development but not pluripotent gene expression. Reproduction 150:1–10

    Article  CAS  Google Scholar 

  • Moro LN, Jarazo J, Buemo C, Hiriart MI, Sestelo A, Salamone DF (2015b) Tiger, Bengal and domestic cat embryos produced by homospecific and interspecific zona-free nuclear transfer. Reprod Domest Anim 50:849–857

    Article  CAS  Google Scholar 

  • Murmann AE, Mincheva A, Scheuermann MO, Gautier M, Yang F, Buitkamp J, Strissel PL, Strick R, Rowley JD, Lichter P (2008) Comparative gene mapping in cattle, Indian muntjac, and Chinese muntjac by fluorescence in situ hybridization. Genetica 134:345–351

    Article  CAS  Google Scholar 

  • Nanda PK, Swain P, Nayak SK, Behera T, Dhama K (2014) Comparative study on enzymatic and explant method in establishing primary culture from different cultivable cells of Indian major carp, Cirrhinus mrigala. Asian J Anim Vet Adv 9:281–291

    Article  Google Scholar 

  • Narbonne P, Miyamoto K, Gurdon J (2012) Reprogramming and development in nuclear transfer embryos and in interspecific systems. Curr Opin Genet Dev 22:450–458

    Article  CAS  Google Scholar 

  • Ogura A, Inoue K, Wakayama T (2013) Recent advancements in cloning by somatic cell nuclear transfer. Philos Trans R Soc B Biol Sci 368:20110329

    Article  Google Scholar 

  • Santos MLT, Borges AA, Neta LBQ, Santos MVO, Oliveira MF, Silva AR, Pereira AF (2016) In vitro culture of somatic cells derived from ear tissue of collared peccary (Pecari tajacu Linnaeus, 1758) in medium with different requirements. Pesqui Vet Bras 36:1194–1202

    Article  Google Scholar 

  • Siengdee P, Klinhom S, Thitaram C, Nganvongpanit K (2018) Isolation and culture of primary adult skin fibroblasts from the Asian elephant (Elephas maximus). PeerJ 6:e4302

    Article  Google Scholar 

  • Simon D (1984) Conservation of animal genetic resources - a review. Livest Prod Sci 11:23–36

    Article  Google Scholar 

  • Srisodsuk S, Duengkae P, Kongprom U, Siriaroonrat B, Duangjai S (2018) The complete mitochondrial genome of Fea’s muntjac (Muntiacus feae Thomas and Doria, 1889) with phylogenetic analysis. Mitochondrial DNA Part B 3:982–983

    Article  Google Scholar 

  • Uemori T, Asada K, Kato I, Harasawa R (1992) Amplification of the 16S-23S spacer region in rRNA operons of mycoplasmas by the polymerase chain reaction. Syst Appl Microbiol 15:181–186

    Article  CAS  Google Scholar 

  • Verma R, Holland MK, Temple-Smith P, Verma PJ (2012) Inducing pluripotency in somatic cells from the snow leopard (Panthera uncia), an endangered felid. Theriogenology 77:220–228 228.e1-2

    Article  CAS  Google Scholar 

  • Volokhov DV, Graham LJ, Brorson KA, Chizhikov VE (2011) Mycoplasma testing of cell substrates and biologics: review of alternative non-microbiological techniques. Mol Cell Probes 25:69–77

    Article  CAS  Google Scholar 

  • Wang W, Lan H (2000) Rapid and parallel chromosomal number reductions in Muntjac deer inferred from mitochondrial DNA phylogeny. Mol Biol Evol 17:1326–1333

    Article  CAS  Google Scholar 

  • Webb SJ, Zychowski GV, Bauman SW, Higgins BM, Raudsepp T, Gollahon LS, Wooten KJ, Cole JM, Godard-Codding C (2014) Establishment, characterization, and toxicological application of loggerhead sea turtle (Caretta caretta) primary skin fibroblast cell cultures. Environ Sci Technol 48:14728–14737

    Article  CAS  Google Scholar 

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Correspondence to Jinpu Wei.

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All procedures in this study involving animals were in accordance with the ethical principles and guidelines of the institution. This study was approved by the Bioethics Committee of China National GeneBank (NO. CNGB-IRB1901-2).

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The authors declare that they have no conflict of interest.

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Editor: Tetsuji Okamoto

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Wang, T., Li, Z., Zheng, D. et al. Establishment and characterization of a fibroblast cell line from postmortem skin of an adult Chinese muntjac (Muntiacus reevesi). In Vitro Cell.Dev.Biol.-Animal 56, 97–102 (2020). https://doi.org/10.1007/s11626-019-00422-8

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