Displaying publications 1 - 20 of 38 in total

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  1. A prospective study of mycobacterial viability in refrigerated, unpreserved sputum batched for up to 8 weeks
    Rashid Ali MR, Parameswaran U, William T, Bird E, Wilkes CS, Lee WK, et al.
    Int J Tuberc Lung Dis, 2015 May;19(5):620-1.
    PMID: 25868033 DOI: 10.5588/ijtld.14.0938
    Matched MeSH terms: Cryopreservation/methods*
  2. Alpha-linolenic acid supplementation in tris extender can improve frozen-thawed bull semen quality
    Kaka A, Wahid H, Rosnina Y, Yimer N, Khumran AM, Behan AA, et al.
    Reprod. Domest. Anim., 2015 Feb;50(1):29-33.
    PMID: 25366298 DOI: 10.1111/rda.12445
    The study was conducted to evaluate the effects of α-linolenic acid (ALA) on frozen-thawed quality and fatty acid composition of bull sperm. For that, twenty-four ejaculates obtained from three bulls were diluted in a Tris extender containing 0 (control), 3, 5, 10 and 15 ng/ml of ALA. Extended semen was incubated at 37°C for 15 min, to allow absorption of ALA by sperm cell membrane. The sample was chilled for 2 h, packed into 0.25-ml straws and frozen in liquid nitrogen for 24 h. Subsequently, straws were thawed and evaluated for total sperm motility (computer-assisted semen analysis), membrane functional integrity (hypo-osmotic swelling test), viability (eosin-nigrosin), fatty acid composition (gas chromatography) and lipid peroxidation (thiobarbituric acid-reactive substances (TBARS)). A higher (p < 0.05) percentage of total sperm motility was observed in ALA groups 5 ng/ml (47.74 ± 07) and 10 ng/ml (44.90 ± 0.7) in comparison with control (34.53 ± 3.0), 3 ng/ml (34.40 ± 2.6) and 15 ng/ml (34.60 ± 2.9). Still, the 5 ng/ml ALA group presented a higher (p < 0.05) percentage of viable sperms (74.13 ± 0.8) and sperms with intact membrane (74.46 ± 09) than all other experimental groups. ALA concentration and lipid peroxidation in post-thawed sperm was higher in all treated groups when compared to the control group. As such, the addition of 5 ng/ml of ALA to Tris extender improved quality of frozen-thawed bull spermatozoa.
    Matched MeSH terms: Cryopreservation/methods
  3. Biobanking of Human Mesenchymal Stem Cells: Future Strategy to Facilitate Clinical Applications
    Yong KW, Choi JR, Wan Safwani WK
    Adv Exp Med Biol, 2016;951:99-110.
    PMID: 27837557
    Human mesenchymal stem cells (hMSCs), a type of adult stem cells that hold great potential in clinical applications (e.g., regenerative medicine and cell-based therapy) due to their ability to differentiate into multiple types of specialized cells and secrete soluble factors which can initiate tissue repair and regulate immune response. hMSCs need to be expanded in vitro or cryopreserved to obtain sufficient cell numbers required for clinical applications. However, long-term in vitro culture-expanded hMSCs may raise some biosafety concerns (e.g., chromosomal abnormality and malignant transformation) and compromised functional properties, limiting their use in clinical applications. To avoid those adverse effects, it is essential to cryopreserve hMSCs at early passage and pool them for off-the-shelf use in clinical applications. However, the existing cryopreservation methods for hMSCs have some notable limitations. To address these limitations, several approaches have to be taken in order to produce healthy and efficacious cryopreserved hMSCs for clinical trials, which remains challenging to date. Therefore, a noteworthy amount of resources has been utilized in research in optimization of the cryopreservation methods, development of freezing devices, and formulation of cryopreservation media to ensure that hMSCs maintain their therapeutic characteristics without raising biosafety concerns following cryopreservation. Biobanking of hMSCs would be a crucial strategy to facilitate clinical applications in the future.
    Matched MeSH terms: Cryopreservation/methods*
  4. Comparative cellular and molecular analyses of pooled bone marrow multipotent mesenchymal stromal cells during continuous passaging and after successive cryopreservation
    Mamidi MK, Nathan KG, Singh G, Thrichelvam ST, Mohd Yusof NA, Fakharuzi NA, et al.
    J Cell Biochem, 2012 Oct;113(10):3153-64.
    PMID: 22615164 DOI: 10.1002/jcb.24193
    The clinical application of human bone marrow derived multipotent mesenchymal stromal cells (MSC) requires expansion, cryopreservation, and transportation from the laboratory to the site of cell implantation. The cryopreservation and thawing process of MSCs may have important effects on the viability, growth characteristics and functionality of these cells both in vitro and in vivo. More importantly, MSCs after two rounds of cryopreservation have not been as well characterized as fresh MSCs from the transplantation perspective. The objective of this study was to determine if the effect of successive cryopreservation of pooled MSCs during the exponential growth phase could impair their morphology, phenotype, gene expression, and differentiation capabilities. MSCs cryopreserved at passage 3 (cell bank) were thawed and expanded up to passage 4 and cryopreserved for the second time. These cells (passive) were then thawed and cultured up to passage 6, and, at each passage MSCs were characterized. As control, pooled passage 3 cells (active) after one round of cryopreservation were taken all the way to passage 6 without cryopreservation. We determined the growth rate of MSCs for both culture conditions in terms of population doubling number (PDN) and population doubling time (PDT). Gene expression profiles for pluripotency markers and tissue specific markers corresponding to neuroectoderm, mesoderm and endoderm lineages were also analyzed for active and passive cultures of MSC. The results show that in both culture conditions, MSCs exhibited similar growth properties, phenotypes and gene expression patterns as well as similar differentiation potential to osteo-, chondro-, and adipo-lineages in vitro. To conclude, it appears that successive or multiple rounds of cryopreservation of MSCs did not alter the fundamental characteristics of these cells and may be used for clinical therapy.
    Matched MeSH terms: Cryopreservation/methods*
  5. Comparative characteristics of spermatozoa harvested and cryopreserved in culture and cryoprotectant media with or without donor serum proteins
    Ata'Allah GA, Adenan NAM, Razali N, Palaniappan K, Saad R, Idris SK, et al.
    Reprod Biol, 2017 Jun;17(2):172-179.
    PMID: 28511996 DOI: 10.1016/j.repbio.2017.04.004
    The objectives of this study is to evaluate the efficacy of protein-free media in the preparation, holding and crypreservation of spermatazoa for use in ART. Normozoospermic semen samples (N=71) were used to compare the effects of media on the survival and quality of spermatozoa when washed and cultured with different media with and without added proteins at 4°C, 15°C, 22°C and 37°C for 0, 4-7 and 24h. Survival and quality of spermatozoa were assessed after freeze-thaw with synthetic cryoprotectant with and without proteins. Ethics/IRB approval was obtained (Ref. 1073.52). Spermatozoa parameters were similar in all media after washing and culture for 24h. Post-thaw survival and quality of spermatozoa was not significantly different 24h after thawing of samples frozen in all cryoprotectant medium. In conclusion synthetic protein-free culture and cryoprotectant media are equal in efficacy to protein-containing media in culture and cryopreservation of spermatozoa . Use of these synthetic media are anticipated to significantly reduce the risk, potentially associated with conventional protein-containing media, of transmission of disease and possibly harmful undeclared proteins to the patient, baby and the healtcare worker. Synthetic media also ensure consistency of quality between batches of media.
    Matched MeSH terms: Cryopreservation/methods*
  6. Fulltext Comparison between mechanical freezer and conventional freezing using liquid nitrogen in normozoospermia
    Rahana AR, Ng SP, Leong CF, Rahimah MD
    Singapore Med J, 2011 Oct;52(10):734-7.
    PMID: 22009393
    INTRODUCTION: This study evaluated the effect of human semen cryopreservation using an ultra-low temperature technique with a mechanical freezer at -85°C as an alternative method to the conventional liquid nitrogen technique at -196°C.
    METHODS: This was a prospective experimental study conducted in the Medically Assisted Conception unit, Department of Obstetrics and Gynaecology, National University Hospital, Malaysia from January 1, 2006 to April 30, 2007. All normozoospermic semen samples were included in the study. The concentration, motility and percentage of intact DNA of each semen sample were assessed before and after freezing and thawing on Days 7 and 30 post freezing.
    RESULTS: Sperm cryopreservation at -85°C was comparable to the conventional liquid nitrogen technique for a period of up to 30 days in a normozoospermic sample. There was no statistical difference in concentration (Day 7 p-value is 0.1, Day 30 p-value is 0.2), motility (Day 7 p-value is 0.9, Day 30 p-value is 0.5) and proportion of intact DNA (Day 7 p-value is 0.1, Day 30 p-value is 0.2) between the ultra-low temperature technique and conventional liquid nitrogen cryopreservation at Days 7 and 30 post thawing.
    CONCLUSION: This study clearly demonstrates that short-term storage of sperm at -85°C could be a viable alternative to conventional liquid nitrogen cryopreservation at -196°C due to their comparable post-thaw results.
    Matched MeSH terms: Cryopreservation/methods*
  7. Cryopreservation of Brassidium Shooting Star orchid using the PVS3 method supported with preliminary histological analysis
    Mubbarakh SA, Rahmah S, Rahman ZA, Sah NN, Subramaniam S
    Appl Biochem Biotechnol, 2014 Jan;172(2):1131-45.
    PMID: 24146369 DOI: 10.1007/s12010-013-0597-0
    Cryopreservation is an alternative, safe, and cost-effective method for long-term plant genetic resource conservation. This study was conducted to optimize the conditions for cryopreserving the protocorm-like bodies (PLBs) of Brassidium Shooting Star orchid with the PVS3 vitrification method. Five parameters were assessed in this study: PLB size, sucrose concentration, preculture duration, PVS3 duration, and unloading duration. The viability of the cryopreserved PLBs was determined using the triphenytetrazolium chloride assay and growth recovery assessments. The optimum condition for the cryopreservation of the PLBs of Brassidium Shooting Star orchid is based on the size range between 3 and 4 mm precultured with half-strength semi-solid MS media supplemented with 0.25 M sucrose for 24 h, followed by treatment with loading solution mixture of 2 M glycerol and 0.4 M sucrose supplemented with half-strength liquid MS media at 25 °C for 20 min. The PLBs were then dehydrated with PVS3 at 0 °C for 20 min prior to immersion in liquid nitrogen; finally, the PLBs were immersed with half-strength liquid MS media supplemented with 1.2 M sucrose for 30 min. Histological analyses displayed denser cytoplasm and voluminous nucleus in the cryopreserved PLBs of Brassidium Shooting Star orchid.
    Matched MeSH terms: Cryopreservation/methods*
  8. Cryopreservation of Citrus aurantifolia seeds and embryonic axes using a desiccation protocol
    Cho EG, Noor NM, Kim HH, Rao VR, Engelmann F
    Cryo Letters, 2002 Sep-Oct;23(5):309-16.
    PMID: 12447490
    The desiccation and freezing tolerance of seeds, with and without testas, and embryonic axes of Citrus aurantifolia were investigated. Seeds were desiccated with silica gel, under the laminar air flow cabinet or by placing them on a laboratory bench. Whatever the desiccation method employed, survival before and after cryopreservation was higher for seeds without testas. When freezing intact seeds, the highest survival percentage (41.3 %) was achieved after desiccation to 7.3 % moisture content (fresh weight basis) on the laboratory bench. Survival of seeds cryopreserved without testas could reach up to 85 % after desiccation under the laminar air flow cabinet or on the laboratory bench, corresponding to moisture contents of 7.1 and 4.5 %, respectively. After desiccation with silica gel, survival reached a maximum of 60.0 %, for a seed moisture content of 3.3 %. Survival of control embryonic axes was high (80-100 %) whatever the sucrose concentration in the preculture medium and the duration of the desiccation period. After cryopreservation, no survival was noted with embryonic axes, which had not been precultured nor desiccated. Survival of non-desiccated embryonic axes after cryopreservation increased progressively in line with increasing sucrose concentrations in the preculture medium, from 7.5 % with 0.1 M sucrose to 77.5 % with 0.7 M sucrose. Survival of desiccated and cryopreserved embryos was always high, whatever the preculture treatment and desiccation period, ranging from 55.8 % to 92.5 %.
    Matched MeSH terms: Cryopreservation/methods*
  9. Cryopreservation of Citrus madurensis embryonic axes by vitrification: importance of loading and treatment with vitrification solution
    Cho EG, Hor YL, Kim HH, Rao VR, Engelmann F
    Cryo Letters, 2002 Sep-Oct;23(5):317-24.
    PMID: 12447491
    This paper investigates the importance of loading and treatment with a vitrification solution on the survival of Citrus madurensis embryonic axes cryopreserved using a vitrification protocol. Among the seven different loading solutions tested, the solution containing 2 M glycerol + 0.4 M sucrose was the most efficient. Of the six vitrification solutions tested, the PVS2 vitrification solution, applied for 20 min at 25 degree C or for 60 min at 0 degree C, ensured the highest survival. A three-step vitrification protocol, involving the treatment of embryonic axes at 0 degree C with half strength PVS2 solution for 20 min then with full strength PVS2 for an additional 40 min was more efficient than a two-step protocol that involved treatment of axes directly with full strength PVS2 solution for 60 min. After rapid immersion in liquid nitrogen, rapid rewarming, unloading in a 1.2 M sucrose solution for 20 min, culture on solid medium with 0.3 M sucrose for 1 day and growth recovery for 4 weeks on standard medium, survival of C. madurensis embryonic axes reached 85 % following the three-step process, compared with 70 % for the two-step process.
    Matched MeSH terms: Cryopreservation/methods*
  10. Cryopreservation of Citrus madurensis embryonic axes encapsulation-dehydration
    Cho EG, Hor YL, Kim HH, Rao VR, Engelmann F
    Cryo Letters, 2002 Sep-Oct;23(5):325-32.
    PMID: 12447492
    In this paper, we demonstrate that C. madurensis embryonic axes can withstand cryopreservation using the encapsulation-dehydration technique. Up to 57.5 % survival was achieved using a standard encapsulation-dehydration protocol, which included pregrowth of encapsulated axes for 16 h in medium containing 0.8 M sucrose + 1 M glycerol, desiccation of beads to around 30 % moisture content (fresh weight basis) followed by rapid freezing. A slightly higher survival percentage (65 %) was obtained using a modified encapsulation-dehydration protocol, which included pretreatment of axes with 2 M glycerol + 0.6 M sucrose for 1 h, concomitantly with their encapsulation in 3 % calcium alginate beads, followed by desiccation of the beads to around 30 % moisture content.
    Matched MeSH terms: Cryopreservation/methods*
  11. Cryopreservation of Citrus madurensis zygotic embryonic axes by vitrification: importance of pregrowth and preculture conditions
    Cho EG, Hor YL, Kim HH, Rao VR, Engelmann F
    Cryo Letters, 2001 Nov-Dec;22(6):391-6.
    PMID: 11788881
    The role of pregrowth and preculture treatments in terms of both medium composition and exposure duration on survival of embryonic axes of Citrus madurensis after cryopreservation using the vitrification procedure was investigated. The optimal pregrowth treatment for excised embryonic axes was a 3-day treatment with 0.1M sucrose. Preculture was also essential in increasing survival after cryopreservation. Among the various media and treatment durations evaluated, a 24h-preculture of embryonic axes on medium with 0.3M sucrose and 0.5M glycerol was found to be optimal. Using these pregrowth and preculture conditions followed by treatment at 25 degrees C for 20 min each with a loading solution (0.4M sucrose + 2.0M glycerol) and then the PVS2 vitrification solution, direct immersion in liquid nitrogen, rapid rewarming, unloading in a 1.2M sucrose solution for 20 min and transfer of embryonic axes on recovery medium, 82.5% survival and regrowth without intermediary callus formation were obtained with C. madurensis embryonic axes.
    Matched MeSH terms: Cryopreservation/methods*
  12. Cryopreservation of Garcinia cowa shoot tips by vitrification: the effects of sucrose preculture and loading treatment on ultrastructural changes in meristematic cells
    Yap LV, Noor NM, Clyde MM, Chin HF
    Cryo Letters, 2011 May-Jun;32(3):188-96.
    PMID: 21766148
    The effects of sucrose preculture duration and loading treatment on tolerance of Garcinia cowa shoot tips to cryopreservation using the PVS2 vitrification solution were investigated. Ultrastructural changes in meristematic cells at the end of the preculture and loading steps were followed in an attempt to understand the effects of these treatments on structural changes in cell membranes and organelles. Increasing preculture duration on 0.3 M sucrose medium from 0 to 3 days enhanced tolerance to PVS2 solution from 5.6 percent (no preculture) to 49.2 percent (3-day preculture). However, no survival was observed after cryopreservation. Examination of meristematic cells by transmission electron microscopy revealed the progressive accumulation of an electron-dense substance in line with increasing exposure durations to 0.3 M sucrose preculture. Treatment with a loading solution (2 M glycerol + 0.4 M sucrose) decreased tolerance of shoot tips to PVS2 vitrification solution and had a deleterious effect on the ultrastructure of G. cowa meristematic cells. This study suggests that G. cowa meristematic cells may lose their structural integrity due to exposure to glycerol present in the loading solution at a 2 M concentration, either due to its high osmotic potential, or due to its cytotoxicity.
    Matched MeSH terms: Cryopreservation/methods*
  13. Cryopreservation of Human Mesenchymal Stem Cells for Clinical Applications: Current Methods and Challenges
    Yong KW, Wan Safwani WK, Xu F, Wan Abas WA, Choi JR, Pingguan-Murphy B
    Biopreserv Biobank, 2015 Aug;13(4):231-9.
    PMID: 26280501 DOI: 10.1089/bio.2014.0104
    Mesenchymal stem cells (MSCs) hold many advantages over embryonic stem cells (ESCs) and other somatic cells in clinical applications. MSCs are multipotent cells with strong immunosuppressive properties. They can be harvested from various locations in the human body (e.g., bone marrow and adipose tissues). Cryopreservation represents an efficient method for the preservation and pooling of MSCs, to obtain the cell counts required for clinical applications, such as cell-based therapies and regenerative medicine. Upon cryopreservation, it is important to preserve MSCs functional properties including immunomodulatory properties and multilineage differentiation ability. Further, a biosafety evaluation of cryopreserved MSCs is essential prior to their clinical applications. However, the existing cryopreservation methods for MSCs are associated with notable limitations, leading to a need for new or improved methods to be established for a more efficient application of cryopreserved MSCs in stem cell-based therapies. We review the important parameters for cryopreservation of MSCs and the existing cryopreservation methods for MSCs. Further, we also discuss the challenges to be addressed in order to preserve MSCs effectively for clinical applications.
    Matched MeSH terms: Cryopreservation/methods*
  14. Cryopreservation of scalps of Malaysian bananas using a pre-growth method
    Sipen P, Anthony P, Davey MR
    Cryo Letters, 2011 May-Jun;32(3):197-205.
    PMID: 21766149
    The effect of preculture with different sugars and mannitol on cryopreservation of scalps of the banana (Musa) cvs. Pisang Mas, Pisang Nangka, Pisang Berangan and Pisang Awak was investigated. Scalps (0.3 square cm) were precultured on semi-solid MS-based medium, containing 0.4 or 0.5 M sucrose, glucose, fructose, trehalose or mannitol, for 14 days under a 16 h light and 8 h dark photoperiod prior to rapid cooling and storage in liquid nitrogen. Explants were rewarmed rapidly in a water bath at 40 degree C for 1 min, followed by recovery on two layers of sterile filter paper overlaying 25 ml aliquots of semi-solid MS-based medium with 5 mg per liter benzylaminopurine, 0.2 mg per liter indole acetic acid and 10 mg per liter ascorbic acid (PM8 medium) for 2 days in the dark. Subsequently, scalps were transferred onto 25 ml aliquots of semi-solid PM8 medium and incubated in the dark for 1 week prior to incubation in the light. Shoot regeneration from 5 - 48 percent of cryopreserved scalps of all the banana cvs., was observed only following preculture with 0.4 or 0.5 M glucose or fructose, and with 0.4 M trehalose for the cvs. Pisang Berangan and Pisang Awak. Preculture with 0.4 M glucose resulted in maximum shoot regeneration of cryopreserved scalps of 10 percent, 13 percent, 42 percent and 48 percent for the cvs. Pisang Mas, Pisang Nangka, Pisang Berangan and Pisang Awak, respectively. Concentrations of 0.5 M trehalose, or 0.4 and 0.5 M sucrose or mannitol were extremely toxic to scalps of all the cvs. investigated.
    Matched MeSH terms: Cryopreservation/methods*
  15. Cryopreservation of shoot tips of recalcitrant and tropical species: Advances and strategies
    Normah MN, Sulong N, Reed BM
    Cryobiology, 2019 04;87:1-14.
    PMID: 30677412 DOI: 10.1016/j.cryobiol.2019.01.008
    There is a pressing need for practical and successful conservation efforts to establish long-term germplasm collections of recalcitrant and tropical species, given the challenge and threat that these plants are facing. Cryopreservation is the only way of conserving some of these species, especially those with temperature or desiccation sensitive (recalcitrant) seeds. This review covers reports on cryopreservation studies of shoot tips (apical and axillary) of tropical and subtropical plants. Since many of these species have recalcitrant seeds, the cryopreservation successes, failures and problems involved with these seeds are also discussed. The methodologies, important factors and steps involved in successful cryopreservation protocols are analyzed. Finally strategies are suggested to develop a successful cryopreservation protocol for new plant species, in particular those with tropical recalcitrant seeds.
    Matched MeSH terms: Cryopreservation/methods*
  16. Cryopreservation of whole seeds and excised embryonic axes of Citrus suhuiensis cv. limau langkat in accordance to their desiccation sensitivity
    Makeen MA, Noor NM, Dussert S, Clyde MM
    Cryo Letters, 2005 Jul-Aug;26(4):259-68.
    PMID: 19827255
    Following the investigation of desiccation sensitivity and freezing tolerance of the whole seed of Citrus suhuiensis cv. limau langkat, desiccation sensitivity and cryopreservation of the excised embryonic axes from the seeds of the same species were examined. Three drying conditions were employed: desiccation by equilibrium for the whole seeds and desiccation in laminar airflow and over silica gel for the excised embryonic axes. The relevance of desiccation sensitivity (WC50) to cryopreservation of whole seeds and excised axes was investigated. High desiccation tolerance (WC50 = 0.034 g H2O x g(-1)dw) was acquired for axes desiccated with faster dehydration rate (1.5 g x g(-1) x h(-1)) in laminar airflow compared to substantially lower desiccation tolerance (WC50 = 0.132 and 0.110 g H2O x g(-1)dw) acquired under slower dehydration rates (1.0 and 0.005 g x g(-1) x h(-1)) for axes desiccated over silica gel and whole seeds desiccated by equilibrium respectively. While few whole seeds (8.3%) survived freezing, high recovery percentages of axes (83.3% and 62.2%) after freezing were obtained under laminar airflow and silica gel drying conditions respectively. Irrespective of the drying method employed, axes survival percentages after exposure to LN temperature commensurate with the desiccation sensitivity pattern. For the whole seeds, a factor other than desiccation sensitivity that limits the tolerance to exposure to LN temperature seems to exist and still needs to be defined.
    Matched MeSH terms: Cryopreservation/methods*
  17. Cryopreserved mesenchymal stromal cell treatment is safe and feasible for severe dilated ischemic cardiomyopathy
    Chin SP, Poey AC, Wong CY, Chang SK, Teh W, Mohr TJ, et al.
    Cytotherapy, 2010;12(1):31-7.
    PMID: 19878080 DOI: 10.3109/14653240903313966
    Bone marrow (BM) mesenchymal stromal cells (MSC) represent a novel therapy for severe heart failure with extensive myocardial scarring, especially when performed concurrently with conventional revascularization. However, stem cells are difficult to transport in culture media without risk of contamination, infection and reduced viability. We tested the feasibility and safety of off-site MSC culture and expansion with freeze-controlled cryopreservation and subsequent rapid thawing of cells immediately prior to implantation to treat severe dilated ischemic cardiomyopathy.
    Matched MeSH terms: Cryopreservation/methods*
  18. Desiccation sensitivity and cryopreservation of excised embryonic axes of Citrus suhuiensis cv. limau madu, Citrumelo [Citrus paradisi macf. × Poncirus trifoliata (l.) raf.] and Fortunella polyandra
    Al Zoubi OM, Normah MN
    Cryo Letters, 2012 May-Jun;33(3):241-51.
    PMID: 22825791
    Excised embryonic axes from seeds of three taxa, namely, Citrus suhuiensis cv. limau madu, Citrumelo (Citrus paradisi x Poncirus trifoliate) and Fortunella polyandra, were desiccated in a laminar airflow, over silica gel, and ultra-rapidly. Desiccation sensitivity (WC50) was estimated for each taxon using the quantal response model. High desiccation tolerance (WC50 = 0.11 g water per g dry mass. g/gdw) was observed for limau madu embryonic axes desiccated in a laminar airflow and ultra-rapidly (WC50 =0.10 g/gdw). Desiccation tolerance was substantially lower (WC50 = 0.19 g/gdw) for silica gel dehydration. Similarly, high desiccation tolerance (WC50 = 0.15 g/gdw) was associated with F. polyandra embryonic axes when desiccated in a laminar airflow, while a lower desiccation tolerance (WC50 = 0.17 g/gdw) was observed with silica gel dehydration. Ultra-rapid desiccation led to the highest desiccation tolerance (WC50 = 0.14 g/gdw). The dehydration rate, however, had no influence on desiccation tolerance (WC50 ~ 0.14 g/gdw) for Citrumelo embryonic axes. After each desiccation period, embryonic axes were directly immersed in liquid nitrogen (LN) followed by rapid rewarming. Normal seedling recovery of 80 to 83% for excised embryonic axes of limau madu was observed for laminar airflow and ultra-rapid dehydration, but for silica gel dehydration, 57% recovery was obtained. Similarly, for Citrumelo, high recoveries of 100% and 97% were obtained from axes desiccated in a laminar airflow and using ultra-rapid dehydration, respectively, whereas a lower value was associated with silica gel dehydration (80%). For F. polyandra, 50% recovery was obtained both for laminar airflow and ultra-rapid dehydration, while much lower recovery (43%) was associated with silica gel dehydration. Regardless of the drying method employed, axis survival percentages following exposure to LN were commensurate with the desiccation sensitivity pattern.
    Matched MeSH terms: Cryopreservation/methods*
  19. Development of a PVS2 droplet-vitrification cryopreservation technique for Aranda Broga Blue orchid protocorm-like bodies (PLBs)
    Ping KS, Poobathy RR, Zakaria R, Subramaniam S
    Cryo Letters, 2018 5 8;38(4):290-298.
    PMID: 29734430
    　　BACKGROUND: Conservation of commercially important ornamental plants is important to maintain its unique beauty to cater the market demands.

    OBJECTIVE: The main objective is to develop an efficient cryopreservation technique for Aranda Broga Blue orchid PLBs using droplet-vitrification method.

    MATERIALS AND METHODS: Several critical factors in cryopreservation were accessed such as preculture concentrations and durations, choice of vitrification solutions, two-step or three-step vitrification, growth recovery medium and PVS2 exposure duration.

    RESULTS: The best growth regeneration percentage (5%) was obtained when 3-4mm PLBs were precultured in 0.2M sucrose for 3 days, followed by osmoprotection for 20 minutes, dehydration in PVS2 for 20 minutes at 0 degree C, LN storage, thawed and unloading for 20 minutes, and growth regeneration in VW10 medium. PLBs were found to be very sensitive to osmotic stress imposed by high molecular weight cryoprotectant such as sucrose and glycerol. Osmotic potential of growth recovery medium is one of the main factors that affect growth recovery in cryopreserved PLBs.

    CONCLUSION: Current report showed possibilities in cryopreserving Aranda Broga Blue PLBs using droplet-vitrification technique. However, further improvement of growth recovery can be done by focussing on approaches that facilitate sufficient water removal from PLBs without causing severe osmotic injuries to the plant cells.

    Matched MeSH terms: Cryopreservation/methods*
  20. Effect of type and concentration of antioxidant on sperm motility, viability, and DNA integrity of climbing perch Anabas testudineus Bloch, 1792 (Pisces: Anabantidae) post-cryopreservation
    Maulida S, Eriani K, Fadli N, Siti-Azizah MN, Kocabas FK, Kocabas M, et al.
    Cryobiology, 2024 Mar;114:104851.
    PMID: 38237749 DOI: 10.1016/j.cryobiol.2024.104851
    Sperm quality is preserved through the crucial involvement of antioxidants, which play a vital role in minimizing the occurrence of reactive oxygen species (ROS) during the cryopreservation process. The suitability of the type and concentration of antioxidants are species-dependent, and this study is crucial in order to improve the quality of the climbing perch sperm post-cryopreservation. Therefore, this study aimed to determine the best type and concentration of antioxidants for cryopreservation of climbing perch Anabas testudineus sperm. To achieve this, 6 types of antioxidants, namely, ascorbic acid, beta-carotene, glutathione, butylated hydroxytoluene (BHT), myo-inositol, and alpha-tocopherol, with inclusion of a control were tested in 3 replications at three concentration levels of 0 mg/L (control), 20 mg/L, 40 mg/L, and 60 mg/L. Sperm was diluted in a glucose-base extender at a ratio of 1:60 (sperm: glucose base), then 10 % DMSO and 5 % egg yolk was added before cryopreservation for two weeks. The results showed that the type and concentration of antioxidants had a significant effect on the motility and viability of cryopreserved climbing perch sperm (P 
    Matched MeSH terms: Cryopreservation/methods
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