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A comparative study on efficiency of adult fibroblasts and amniotic fluid-derived stem cells as donor cells for production of hand-made cloned buffalo (Bubalus bubalis) embryos

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Abstract

The efficiency of two cell types, namely adult fibroblasts, and amniotic fluid stem (AFS) cells as nuclear donor cells for somatic cell nuclear transfer by hand-made cloning in buffalo (Bubalus bubalis) was compared. The in vitro expanded buffalo adult fibroblast cells showed a typical “S” shape growth curve with a doubling time of 40.8 h and stained positive for vimentin. The in vitro cultured undifferentiated AFS cells showed a doubling time of 33.2 h and stained positive for alkaline phosphatase, these cells were also found positive for undifferentiated embryonic stem cell markers like OCT-4, NANOG and SOX-2, which accentuate their pluripotent property. Further, when AFS cells were exposed to corresponding induction conditions, these cells differentiated into osteogenic, adipogenic and chondrogenic lineages which was confirmed through alizaran, oil red O and alcian blue staining, respectively. Cultured adult fibroblasts and AFS cells of passages 10–15 and 8–12, respectively, were used as nuclear donors. A total of 94 embryos were reconstructed using adult fibroblast as donor cells with cleavage and blastocyst production rate of 62.8 ± 1.8 and 19.1 ± 1.5, respectively. An overall cleavage and blastocyst formation rate of 71.1 ± 1.2 and 29.9 ± 2.2 was obtained when 97 embryos were reconstructed using AFS cells as donor cells. There were no significant differences (P > 0.05) in reconstructed efficiency between the cloned embryos derived from two donor cells, whereas the results showed that there were significant differences (P < 0.05) in cleavage and blastocyst rates between the cloned embryos derived from two donor cell groups. Average total cell numbers for blastocyst generated using AFS cells (172.4 ± 5.8) was significantly (P < 0.05) higher than from adult fibroblasts (148.2 ± 6.1). This study suggests that the in vitro developmental potential of the cloned embryos derived from AFS cells were higher than that of the cloned embryos derived from adult fibroblasts in buffalo hand-made cloning.

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References

  • Arnhold S, Glüer S, Hartmann K, Raabe O, Addicks K, Wenisch S, Hoopmann M (2011) Amniotic fluid stem cells: growth dynamics and differentiation potential after a CD-117-based selection procedure. Stem Cells Int 2011:715341

    Article  CAS  Google Scholar 

  • Berstein EF, Chen YQ, Kopp JB, Fisher L, Brown DB, Hahn PJ, Robey FA, Lakkakorpi J, Uitto J (1996) Long term sun exposure alters the collagen of the papillary dermis. J Am Acad Dermatol 34:209–218

    Article  Google Scholar 

  • Bhojwani S, Vajta G, Callesen H, Roschlau K, Kuwer A, Becker F, Alm H, Torner H, Kanitz W, Poehland R (2005) Developmental competence of HMC™ derived bovine cloned embryos obtained from somatic nuclear transfer of adult fibroblast and granulosa cells. J Reprod Dev 51:465–475

    Article  Google Scholar 

  • Bossolasco P, Montemurro T, Cova L, Zangrossi S, Calzarossa C, Buiatiotis S, Soligo D, Bosari S, Silani V, Deliliers GL, Rebulla P, Lazzari L (2006) Molecular and phenotypic characterization of human amniotic fluid cells and their differentiation potential. Cell Res 16:329–336

    Article  CAS  Google Scholar 

  • Boyer LA, Lee TI, Cole MF, Johnstone SE, Levine SS, Zucker JP, Guenther MG, Kumar RM, Murray HL, Jenner RG, Gifford DK, Melton DA, Jaenisch R, Young RA (2005) Core transcriptional regulatory circuitry in human embryonic stem cells. Cell 122:947–956

    Article  CAS  Google Scholar 

  • Chauhan MS, Singla SK, Palta P, Manik RS, Tomer OS (1998) Development of in vitro produced buffalo (Bubalus bubalis) embryos in relation to time. Asian Aust J Anim Sci 11:398–403

    Article  Google Scholar 

  • Chen J, Lu Z, Cheng D, Peng S, Wang H (2011) Isolation and characterization of porcine amniotic fluid-derived multipotent stem cells. PLoS One 6:e19964

    Article  CAS  Google Scholar 

  • Cheong HT, Takahashi Y, Kanagawa H (1993) Birth of mice after transplantation of early cell-cycle-stage embryonic nuclei into enucleated oocytes. Biol Reprod 48:958–963

    Article  CAS  Google Scholar 

  • Choi SA, Lee JH, Kim KJ, Kim EY, Park KS, Park YB, Li X, Ha YN, Park JY, Kim MK (2011) Isolation and characterization of mesenchymal stem cells derived from human amniotic fluid. Reprod Fertil Dev 23:243–253

    Article  Google Scholar 

  • De Coppi P, Bartsch G, Siddiqui MM, Xu T, Santos CC, Perin L, Mostoslavsky G, Serre AC, Snyder EY, Yoo JJ, Furth ME, Soker S, Atala A (2007) Isolation of amniotic stem cell lines with potential for therapy. Nat Biotechnol 25:100–106

    Article  Google Scholar 

  • Dev K, Giri SK, Kumar A, Yadav A, Singh B, Gautam SK (2010) Derivation, characterization and differentiation of buffalo (Bubalus bubalis) amniotic fluid derived stem cells. Reprod Dom Anim 47:704–711

    Article  Google Scholar 

  • Fortin S, Mercier LM, Camby I, Spiegl-Kreinecker S, Berger W, Lefranc F (2010) Galectin-1 is implicated in the protein kinase Cε/Vimentin-controlled trafficking of integrin-β1 in glioblastoma cells. Brain Pathol 20:39–49

    Article  CAS  Google Scholar 

  • George A, Sharma R, Singh KP, Panda SK, Singla SK, Palta P, Manik R, Chauhan MS (2011) Production of cloned and transgenic embryos using buffalo (Bubalus bubalis) embryonic stem cell-like cells isolated from in vitro fertilized and cloned blastocysts. Cell Reprogram 13:263–272

    Article  CAS  Google Scholar 

  • Gosden CM (1983) Amniotic fluid cell types and culture. Br Med Bull 39:348–354

    CAS  Google Scholar 

  • Hanson C, Caisander G (2005) Human embryonic stem cells and chromosome stability. APMIS 113:751–755

    Article  Google Scholar 

  • Hart AH, Hartley L, Ibrahim M, Robb L (2004) Identification, cloning and expression analysis of the pluripotency promoting NANOG genes in mouse and human. Dev Dyn 230:187–198

    Article  CAS  Google Scholar 

  • Heyman Y, Zhou Qi, Lebourhis D, Chavatte-Palmer P, Renard JP, Vignon X (2002) Novel approaches and hurdles to somatic cloning in cattle. Cloning Stem Cells 4:47–55

    Article  CAS  Google Scholar 

  • Hiiragi T, Solter D (2005) Reprogramming is essential in nuclear transfer. Molec Repro Dev 70:417–421

    Article  CAS  Google Scholar 

  • Hua J, Yu H, Liu S, Dou Z, Sun Y, Jing X, Yang C, Lei A, Wang H, Gao Z (2009) Derivation and characterization of human embryonic germ cells: serum free culture and differentiation potential. Reprod Biomed Online 19:238–249

    Article  Google Scholar 

  • In’t Anker PS, Scherjon SA, Keur CK, Noort WA, Claas FHJ, Willemze R, Fibbe WE, Kanhai HHH (2003) Amniotic fluid as a novel source of mesenchymal stem cells for therapeutic transplantation. Blood 102:1548–1549

    Article  Google Scholar 

  • In’t Anker PS, Scherjon SA, Keur CK, Groot-Swings GMJS, Claas FHJ, Fibbe WE, Kanhai HHH (2004) Isolation of mesenchymal stem cells of fetal or maternal origin from human placenta. Stem Cells 22:1338–1345

    Article  Google Scholar 

  • Kato Y, Tsunoda Y (2010) Role of donor nulcei in cloning efficiency: can the ooplasm reprogram any nucleus. Int J Dev Biol 54:1623–1629

    Article  Google Scholar 

  • Kato Y, Tani T, Tsunoda Y (2000) Cloning of calves from various somatic cell types of male and female adult, newborn and fetal cows. J Reprod Fertil 120:231–237

    Article  CAS  Google Scholar 

  • Kim J, Lee Y, Kim H, Hwang KJ, Kwon HC, Kim SK, Cho DJ, Kang SG, You J (2007) Human amniotic fluid-derived cells have characteristics of multipotent stem cells. Cell Prolif 40:75–90

    Article  CAS  Google Scholar 

  • Kruip TAM, Dieleman SJ (1982) Macroscopic classification of bovine follicles and its validation by micromorphological and steroid biochemical procedures. Reprod Nutr Dev 22:465–473

    Article  CAS  Google Scholar 

  • Li X, Li Z, Jouneau A, Zhou Q, Renard JP (2003) Nuclear transfer: progress and quandaries. Reprod Biol Endocrinol 1:84

    Article  Google Scholar 

  • Li C, Zhou J, Shi G, Ma Y, Yang Y, Gu J, Yu H, Jin S, Wei Z, Chen F, Jin Y (2009) Pluripotency can be rapidly and efficiently induced in human amniotic fluid-derived cells. Hum Mol Genet 18:4340–4349

    Article  CAS  Google Scholar 

  • Liu CQ, Guo Y, Guan WJ, Ma YH, Zhang HH, Tang XX (2008) Establishment and biological characteristics of luxi cattle fibroblast bank. Tissue Cell 40:417–424

    Article  CAS  Google Scholar 

  • Maitra A, Arking DE, Shivapurkar N, Ikeda M, Stastny V, Kassauei K, Sui G, Cutler DJ, Liu Y, Brimble SN, Noaksson K, Hyllner J, Schulz TC, Zeng X, Freed WJ, Crook J, Abraham S, Colman A, Sartipy P, Matsui S, Carpenter M, Gazdar AF, Rao M, Chakravarti A (2005) Genomic alterations in cultured human embryonic stem cells. Nat Genet 37:1099–1103

    Article  CAS  Google Scholar 

  • Mauro A, Turriani M, Loannoni A, Russo V, Martelli A, Di Giacinto O, Nardinocchi D, Berardinelli P (2010) Isolation, characterization, and in vitro differentiation of ovine amniotic stem cells. Vet Res Commun 34:25–28

    Article  Google Scholar 

  • Miyoshi K, Rzucidlo SJ, Pratt SL, Stice SL (2003) Improvements in cloning efficiencies may be possible by increasing uniformity in recipient oocytes and donor cells. Biol Reprod 68:1079–1086

    Article  CAS  Google Scholar 

  • Mizutani E, Ono T, Li C, Maki-Suetsugu R, Wakayama T (2008) Propagation of senescent mice using nuclear transfer embryonic stem cell lines. Genesis 46:478–483

    Article  Google Scholar 

  • Munoz M, Rodriguez A, De Frutos C, Caamano JN, Diez C, Facal N, Gomez E (2008) Conventional pluripotency markers are unspecific for bovine embryonic-derived cell-lines. Theriogenology 69:1159–1164

    Article  CAS  Google Scholar 

  • Panarace M, Aguero JI, Garrote M, Jauregui G, Segovia A, Cane L, Gutierrez J, Marfil M, Rigali F, Pugliese M (2007) How healthy are clones and their progeny: 5 years of field experience. Theriogenology 67:142–151

    Article  CAS  Google Scholar 

  • Parolini O, Soncini M, Evangelista M, Schmidt D (2009) Amniotic membrane and amniotic fluid-derived cells: potential tools for regenerative medicine. Regen Med 4:275–291

    Article  CAS  Google Scholar 

  • Powell AM, Talbot NC, Wells KD, Kerr DE, Pursel VG, Wall RJ (2004) Cell donor influences success of producing cattle by somatic cell nuclear transfer. Biol Reprod 71:210–216

    Article  CAS  Google Scholar 

  • Priest RE, Marimuthu KM, Priest JH (1978) Origin of cells in human amniotic fluid cultures. Lab Invest 39:106–109

    CAS  Google Scholar 

  • Priya D, Selokar NL, Raja AK, Saini M, Sahare AA, Nala N, Palta P, Chauhan MS, Manik RS, Singla SK (2014) Production of wild buffalo (Bubalus arnee) embryos by interspecies somatic cell nuclear transfer using domestic buffalo (Bubalus bubalis) oocytes. Reprod Domest Anim 49:343–351

    Article  CAS  Google Scholar 

  • Prusa AR, Marton E, Rosner M, Bernaschek G, Hengstschläger M (2003) OCT4-expressing cells in human amniotic fluid: a new source for stem cell research. Hum Reprod 18:1489–1493

    Article  Google Scholar 

  • Prusa AR, Marton E, Rosner M, Bettelheim D, Lubec G, Pollack A, Bernaschek G, Hengstschläger M (2004) Neurogenic cells in human amniotic fluid. Am J Obstet Gynecol 191:309–314

    Article  Google Scholar 

  • Rho GJ, Kumar BM, Balasubramanian SS (2009) Porcine mesenchymal stem cells– current technological status and future perspective. Front Biosci 14:3942–3961

    Article  CAS  Google Scholar 

  • Rideout WM, Wakayama T, Wutz A, Eggan K, Jackson-Grusby L, Dausman J, Yanagimachi R, Jaenisch R (2000) Generation of mice from wild-type and targeted ES cells by nuclear cloning. Nat Genet 24:109–110

    Article  CAS  Google Scholar 

  • Sadeesh EM, Kataria M, Balhara S, Yadav PS (2014) Expression profile of developmentally important genes between hand-made cloned buffalo embryos produced from reprogramming of donor cell with oocytes extract and selection of recipient cytoplast through brilliant cresyl blue staining and in vitro fertilized embryos. J Assist Reprod Genet. doi:10.1007/s10815-014-0316-y

    Google Scholar 

  • Saha A, Panda SK, Chauhan MS, Manik RS, Palta P, Singla SK (2012) Birth of cloned calves from vitrified-warmed zona-free buffalo (Bubalus bubalis) embryos produced by hand-made cloning. Reprod Fertil Dev 25:860–865

    Article  Google Scholar 

  • Saikhun J, Kitiyanant N, Songtaveesin C, Pavasuthipaisit K, Kitiyanant Y (2004) Development of swamp buffalo (Bubalus bubalis) embryos after parthenogenetic activation and nuclear transfer using serum fed or starved fetal fibroblasts. Reprod Nutr Dev 44:65–78

    Article  Google Scholar 

  • Selokar NL, Saini M, Muzaffer M, Krishnakanth G, Saha AP, Manik RS, Chauhan MS, Palta P, Madan P, Singla SK (2012) Roscovitine treatment improves synchronization of donor cell cycle in G0/G1 stage and in vitro development of hand-made cloned buffalo (Bubalus bubalis) embryos. Cell Reprogram 14:146–154

    CAS  Google Scholar 

  • Shah R, George A, Singh MK, Kumar D, Chauhan MS, Manik RS, Palta P, Singla SK (2008) Hand-made cloned buffalo (Bubalus bubalis) embryos: comparison of different media and culture systems. Cloning Stem Cells 10:435–442

    Article  CAS  Google Scholar 

  • Shah RA, George A, Singh MK, Kumar D, Anand T, Chauhan MS, Manik RS, Palta AP, Singla SK (2009) Pregnancies established from hand-made cloned blastocyst recontstuectd using skin fibroblast in buffalo (Bubalus bubalis). Theriogenology 71:1215–1219

    Article  CAS  Google Scholar 

  • Shamblott MJ, Axelman J, Wang S, Bugg EM, Littlefield JW, Donovan PJ, Blumenthal PD, Huggins GR, Gearhart JD (1998) Derivation of pluripotent stem cells from cultured human primordial germ cells. Proc Natl Acad Sci USA 95:13726–13731

    Article  CAS  Google Scholar 

  • Shi D, Lu F, Wei Y, Cui Y, Yang S, Wei J, Liu Q (2007) Buffalos (Bubalus bubalis) cloned by nuclear transfer of somatic cells. Biol Reprod 77:285–291

    Article  CAS  Google Scholar 

  • Soliman MK (1975) Studies on physiological chemistry of allantoic fluid of buffalo at various periods of pregnancy. Indian Vet J 52:106–112

    CAS  Google Scholar 

  • Srirattana K, Lorthongpanich C, Laowtammathro C, Imsoonthornruksa S, Ketudat-Cairns M, Phermthai T, Nagai T, Parnpai R (2010) Effect of donor cell types on developmental potential of cattle (Bos taurus) and swamp buffalo (Bubalus bubalis) cloned embryos. J Reprod Dev 56:49–54

    Article  Google Scholar 

  • Thomson JA, Itskovitz-Eldor J, Shapiro SS, Waknitz MA, Swiergiel JJ, Marshal VS, Jones JM (1998) Embryonic stem cell lines derived from human blastocyst. Science 282:1145–1147

    Article  CAS  Google Scholar 

  • Tsai MS, Lee JL, Chang YJ, Hwang SM (2004) Isolation of human multipotent mesenchymal stem cells from second trimester amniotic fluid using a novel two-stage culture protocol. Hum Reprod 19:1450–1456

    Article  Google Scholar 

  • Vajta G (2007) Hand-made cloning: the future way of nuclear transfer. Trends Biotechnol 25:250–253

    Article  CAS  Google Scholar 

  • Wakayama T, Rodriguez I, Perry AC, Yanagimachi R, Mombaerts P (1999) Mice cloned from embryonic stem cells. Proc Natl Acad Sci USA 96:14984–14989

    Article  CAS  Google Scholar 

  • Wakayama S, Mizutani E, Kishigami S, Thuan NV, Ohta H, Hikichi T, Bui HT, Miyake M, Wakayama T (2005) Mice cloned by nuclear transfer from somatic and ntES cells derived from the same individuals. J Reprod Dev 51:765–772

    Article  Google Scholar 

  • Yadav PS, Mann A, Singh V, Yashveer S, Sharma RK, Singh I (2010) Expression of pluripotency genes in buffalo (Bubalus bubalis) amniotic fluid cells. Reprod Dom Anim 46:705–711

    Article  Google Scholar 

  • Yang X, Smith SL, Tian XC, Lewin HA, Renard JP, Wakayama T (2007) Nuclear reprogramming of cloned embryos and its implications for therapeutic cloning. Nat Genet 39:295–302

    Article  CAS  Google Scholar 

  • Zhao XE, Zheng YM (2010) Development of cloned embryos from porcine neural stem cells and amniotic fluid-derived stem cells. Animal 4:921–929

    Article  CAS  Google Scholar 

  • Zheng YM, Zhao XE, An ZX (2009) Neurogenic differentiation of EGFP gene transfected amniotic fluid derived stem cells from pigs at intermediate and late gestational ages. Reprod Domest Anim 45:78–82

    Google Scholar 

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Acknowledgments

The authors would like to thank Dr. Inderjeet Singh, Director, ICAR-Central Institute for Research on Buffaloes, for providing the necessary facilities for carrying out this work. We are also thankful to Dr. B S Punia for reviewing the manuscript. Funding support from Indian Council of Agricultural Research (ICAR), New Delhi is gratefully acknowledged.

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The authors declare no conflict of interest.

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Authors and Affiliations

  1. Division of Animal Physiology and Reproduction, ICAR-Central Institute for Research on Buffaloes, Hisar, 125001, India

    Sadeesh EM & P. S. Yadav

  2. Division of Biochemistry, ICAR-Indian Veterinary Research Institute, Bareilly, 243122, India

    Meena Kataria

  3. Department of Veterinary Physiology and Biochemistry, Lala Lajpat Rai University of Veterinary and Animal Sciences, Hisar, 125004, India

    Fozia Shah

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  1. Sadeesh EM
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  2. Meena Kataria
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Correspondence to Sadeesh EM.

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EM, S., Kataria, M., Shah, F. et al. A comparative study on efficiency of adult fibroblasts and amniotic fluid-derived stem cells as donor cells for production of hand-made cloned buffalo (Bubalus bubalis) embryos. Cytotechnology 68, 593–608 (2016). https://doi.org/10.1007/s10616-014-9805-1

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