Isolation, culturing and characterization of feeder-independent amniotic fluid stem cells in buffalo (Bubalus bubalis)

Dev, Kapil; Gautam, Sanjeev K.; Giri, Shiv Kumar; Kumar, Anil; Yadav, Anita; Verma, Vinod; Kumar, Parvin; Singh, Birbal

doi:10.1016/j.rvsc.2011.09.007

Cited by 15 publications

(12 citation statements)

References 36 publications

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“…The cells were found to express molecular markers that are specific to pluripotent ES cells. The fetal and AF-derived cells in buffaloes have already been found to express key pluripotency molecular markers Dev et al 2011). In this study in long term cultures cells were able to proliferate up to 75 days and their proliferation rate was more up to 10 passages then decreased later may be due to accumulations of more non-dividing cells, was confirmed by earlier study conducted by Gupta et al (2007) in continuous culture of goat fibroblasts cells.…”

Section: Discussionsupporting

confidence: 86%

“…Among animals, amniotic fluid (AF) derived cells have been reported in rodents (Marcus et al 2008), porcine (Zheng et al 2009) and buffalo . Review of the literature reveals the increasing demand of research on stem cells in livestock assisted reproduction and veterinary applications (Dev et al 2011;Nowak-Imialek et al 2011;Singh et al 2011;Yadav et al 2012). Further, the availability of an easily accessible and reproducible cell source may greatly facilitate the development of stem cell-based tissue engineering and therapies for regenerative veterinary medicine.…”

Section: Introductionmentioning

confidence: 99%

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Culture, characterization and differentiation of cells from buffalo (Bubalus bubalis) amnion

et al. 2012

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Stem cells present an important tool in livestock assisted reproduction and veterinary therapeutic field such as tissue engineering. We report for the first time isolation of pluripotent stem cell-like cells expressing pluripotency markers (alkaline phospahatase, OCT-4, NANOG and SOX-2) from the amnion of water buffalo (Bubalus bubalis). The cells showed no apparent abnormalities in their chromosomal profiles before and after cryopreservation. The cytochemical staining revealed that pluripotent cells were capable of undergoing directed differentiation in vitro into osteocytes. It could be inferred that amnionderived pluripotent stem cell-like cells can be isolated, cultured for many passages and differentiated into mesoderm lineage, and may be an alternative source to mesenchymal stem cells. These cells can have applications in assisted reproduction, developmental biological and regenerative medicine.

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Section: Discussionsupporting

confidence: 86%

Section: Introductionmentioning

confidence: 99%

Culture, characterization and differentiation of cells from buffalo (Bubalus bubalis) amnion

et al. 2012

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“…Dev et al. () reported that the AFCs of buffalo present a pleomorphic shape, including different sizes and shapes, but in culture flasks the pre‐dominant shape was similar to that of fibroblasts. Under scanning microscope, the presence of pentagonal, round and star‐shaped cells was noted, as well as cells in division.…”

Section: Discussionmentioning

confidence: 99%

Effects of Prolonged in vitro Culture and Cryopreservation on Viability, DNA Fragmentation, Chromosome Stability and Ultrastructure of Bovine Cells from Amniotic Fluid and Umbilical Cord

Cunha

Martins

Silva

et al. 2014

Reprod Domestic Animals

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The objective of this work was to study cellular types that did not participated in the gastrulation process, amniotic fluid cells (AFCs) and umbilical cord cells (UCCs), in conditions of long-term culture and cryopreserved with different solutions. The AFCs and UCCs were used in a comparative study with ear fibroblast cells (EFCs) that were cultured in vitro until 20 cellular passages and cryopreserved in 10% dimethylsulphoxide (DMSO), 5% dimethyl formamide (DMF) and 7% glycerol (Gly) solutions. The cellular viability, ultrastructure, DNA fragmentation and chromosome stability were evaluated to determine the cellular type most resistant. In all cell types, it was possible to evaluate the AFCs until 15 passages and UCCs until 20 passages with different periods of cellular growth to reach the confluence phase. Solutions containing 10% DMSO ensured viability of 90.33 ± 5.58%, 90.56 ± 4.40% and 81.90 ± 3.31%, respectively for EFCs, AFCs and UCCs, being significantly more efficient and with less variation than other cryoprotectant solutions. The AFCs were more sensitive to cryopreservation and presented low viability rate at the passage 20 (17.2 ± 8.87%). There was no change in karyotype and nuclear fragmentation was low in all cellular passages studied. With the scanning electron analysis was possible the characterization of AFCs and UCCs in suspension. The three cellular types of cells presented different shapes and characteristics on the surface. The results demonstrate that bovine AFCs and UCCs can be isolated, cultured in vitro and cryopreserved in 10% DMSO, not causing damage to DNA and chromosomes. The UCCs were more resistant than AFCs in all aspects.

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“…The AFS cells were separated by centrifugation (3000 g , 10 min) and washed twice with phosphate buffered saline (PBS). The cells were seeded at density of 103 cells/cm 2 in six‐well culture plates containing cell culture medium (DMEM supplemented with 10% FBS, 1% non‐essential amino acids, 1% penicillin/streptomycin/ampicillin, 1% vitamin solution) and incubated in humidified CO 2 incubator (Lark, China) at 38.5 ± 0.5°C in the presence of 5% CO 2 in humidified air (Dev et al. 2010).…”

Section: Methodsmentioning

confidence: 99%

Derivation, Characterization and Differentiation of Buffalo (Bubalus bubalis) Amniotic Fluid Derived Stem Cells

Dev

Giri

Kumar

et al. 2011

Reprod Domestic Animals

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Amniotic fluid cells (AFCs) are obtained from amnion for pre-natal analysis and can be cultured in vitro. Heterogeneous amniotic fluid (AF) contains various cell types, and it is believed that some of these cells possess the stem cell properties. The aim of this study was to characterize these cells by phenotypical and genotypical means in buffalo. The differentiation potential of amniotic fluid stem (AFS) cells was carried out by converting these cells into neurons. The AFCs were cultured without feeder cells in DMEM containing 16% foetal bovine serum, 1% penicillin/streptomycin and 1%l-glutamine in 5% CO(2) at 38.5 ± 0.5 °C in a CO(2) incubator. After 6 days of culture, different types of cells viz., star shaped (62.7%), spherical without nucleus (1.9%), spherical with nucleus (26.4%), pentagonal (0.4%) and free floating/rounded cells (8.3%) were observed. Most of the cells started anchorage-dependent growth after day 7 of the culture. Expression of Oct-4, Sox-2, Nanog, alkaline phosphatase, 18s rRNA, stem cell factor, cyclin A, Nestin and FGF-5 was observed from the AFS cells in different passages with PCR amplicon of 314, 277, 317, 180, 162, 216, 421, 307 and 210 bp, respectively. During the differentiation step, at day 6, neuron-like cells could be clearly identified and confirmed with Nestin-specific RT-PCR. The cells were found to have a normal karyotype at different passages. These results may contribute towards establishing non-embryonic pluripotent stem cells for various therapeutic and reproductive biotechnological applications in the species.

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Isolation, culturing and characterization of feeder-independent amniotic fluid stem cells in buffalo (Bubalus bubalis)

Cited by 15 publications

References 36 publications

Culture, characterization and differentiation of cells from buffalo (Bubalus bubalis) amnion

Culture, characterization and differentiation of cells from buffalo (Bubalus bubalis) amnion

Effects of Prolonged in vitro Culture and Cryopreservation on Viability, DNA Fragmentation, Chromosome Stability and Ultrastructure of Bovine Cells from Amniotic Fluid and Umbilical Cord

Derivation, Characterization and Differentiation of Buffalo (Bubalus bubalis) Amniotic Fluid Derived Stem Cells

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