Displaying publications 1 - 20 of 93 in total

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  1. Rahmah S, Ahmad Mubbarakh S, Soo Ping K, Subramaniam S
    ScientificWorldJournal, 2015;2015:961793.
    PMID: 25861687 DOI: 10.1155/2015/961793
    Protocorm-like bodies (PLBs) of Brassidium Shooting Star orchid were successfully cryopreserved using droplet-vitrification method. Vitrification based cryopreservation protocol is comprised of preculture, osmoprotection, cryoprotection, cooling, rewarming, and growth recovery and each and every step contributes to the achievement of successful cryopreservation. In order to reveal the lethal and nonlethal damage produced by cryopreservation, histological observation, scanning electron microscopy (SEM), and biochemical analysis were carried out in both cryopreserved and noncryopreserved PLBs of Brassidium Shooting Star orchid comparing with the control PLBs stock culture. Histological and scanning electron microscopy analyses displayed structural changes in cryopreserved PLBs due to the impact of cryoinjury during exposure to liquid nitrogen. Total soluble protein significantly increased throughout the dehydration process and the highest value was achieved when PLBs were stored in liquid nitrogen. Ascorbate peroxidase (APX) and catalase (CAT) showed the highest enzyme activities in both dehydration and cryostorage treatments indicating that stress level of PLBs was high during these stages.
    Matched MeSH terms: Cryopreservation*
  2. Al Zoubi OM, Normah MN
    Cryo Letters, 2015 Nov-Dec;36(6):379-91.
    PMID: 26963884
    To further understand the survival characteristics of desiccation-sensitive excised embryonic axes of Fortunella polyandra to desiccation and cryopreservation it is necessary to study the impact of drying rates on both the ultrastructure and electrolyte leakage.
    Matched MeSH terms: Cryopreservation
  3. Chin AHB, Sandhu S, Caughey L, Ahmad MF, Peate M
    Hum Fertil (Camb), 2023 Dec;26(2):385-397.
    PMID: 37177817 DOI: 10.1080/14647273.2023.2209831
    Upon legalization of social egg freezing in Singapore from 2023 onwards, compulsory pre-procedure counselling is mandated for all prospective patients to enable informed choice about whether to undergo the procedure. Being a newly introduced medical procedure in Singapore, there are currently no clear directives on what pre-procedure counselling for elective egg freezing should entail. Due to pervasive media and internet influences, prospective egg freezing patients could be misled into believing that the procedure represents a guaranteed path to future motherhood, contrary to statements by professional bodies such as the American Society for Reproductive Medicine (ASRM) and the British Fertility Society (BFS). Hence, comprehensive counselling is recommended to provide women with evidence-based information (e.g. success rates of social egg freezing for women of their age) to ensure they make informed decisions and to avoid possible decision regret. For this purpose, a systematic protocol and methodology for pre-procedure counselling of women considering elective egg freezing was developed, incorporating flowcharts and decision trees that are specifically tailored to the unique sociocultural values and legal restrictions in Singapore. Questions relating to the why, what, how, where and when of the egg freezing procedure should be addressed, which could serve as a roadmap to facilitate informed decision-making by women considering elective egg freezing.
    Matched MeSH terms: Cryopreservation/methods
  4. Thiagarajah K, Wong CY, Vijayan VV, Ooi GC, Ng MT, Cheong SK, et al.
    Transfusion, 2015 May;55(5):1028-32.
    PMID: 25472857 DOI: 10.1111/trf.12950
    Processed umbilical cord blood (UCB) must be stored at cryogenic temperature at all times to maintain the quality and viability of the cells. However, a challenge is presented in the form of moving a large number of cryopreserved UCB samples to a new location. In this report, we share our experience on relocating more than 100,000 units of cryopreserved UCB samples stored in 12 liquid nitrogen freezers (LNFs) to our new laboratory.
    Matched MeSH terms: Cryopreservation/methods*
  5. Antony JJ, Mubbarakh SA, Mahmood M, Subramaniam S
    Appl Biochem Biotechnol, 2014 Feb;172(3):1433-44.
    PMID: 24218184 DOI: 10.1007/s12010-013-0636-x
    Histological observation and scanning electron microscopy analyses in Dendrobium Bobby Messina indicates the cellular process of cryopreserved protocorm-like bodies (PLBs) was different comparative to non-cryopreserved PLBs. The cellular process was not only modified by the freezing and thawing effect but also due to the dehydration process itself during the cryopreservation procedure. Histological observation in Dendrobium Bobby Messina in encapsulation-dehydration method indicated that the degree of plasmolysis causes more cellular changes to the cryopreserved PLBs comparative to non-cryopreserved and stock culture PLBs. These results revealed higher amount of homogenous cell population and denser cytoplasm in cryopreserved PLBs. Histological analysis also revealed more voluminous nucleus in cryopreserved PLBs comparative to non-cryopreserved PLBs and PLBs stock culture. In contrast, scanning electron microscope analysis showed severe damages in cryopreserved PLBs and non-cryopreserved PLBs comparative to the PLBs stock culture which in return could be the possible reason of no regrowth in encapsulation-dehydration method. Damages incurred were on top part, side part, and at the stomata of the PLBs. Histological observation and scanning electron microscopy analyses in Dendrobium Bobby Messina indicates that the degree of plasmolysis causes changes in the cellular process of PLBs from cryopreserved PLBs was different comparative to non-cryopreserved PLBs.
    Matched MeSH terms: Cryopreservation*
  6. Suresh K, Init I, Reuel PA, Rajah S, Lokman H, Khairul Anuar A
    Parasitol Res, 1998;84(4):321-2.
    PMID: 9569099
    Matched MeSH terms: Cryopreservation/methods*
  7. Ang HH, Chan KL, Mak JW
    Chemotherapy, 1997 Sep-Oct;43(5):311-5.
    PMID: 9309363 DOI: 10.1159/000239583
    Eleven Malaysian Plasmodium falciparum isolates were cultured in vitro and later subjected to antimalarial evaluations in 96-well microtiter plates. After cryopreservation, the IC50 (nM) for ST 195, ST 196, ST 197, ST 244 and ST 245 isolates were, respectively: 180.9, 198.7, 482.0, 580.0 and 690.1 for chloroquine; 3.4, 3.4, 9.2, 4.0 and 5.8 for mefloquine; 21.9, 10.5, 40.7, 40.1 and 48.7 for quinine; 136.7, 58.8, 116.4, 29.4 and 95.4 for cycloguanil, and 48.3, 57.5, 47.4, 61.5 and 37.8 for pyrimethamine. Before cryopreservation they were 172.5, 141.5, 453.2, 636.0 and 651.6 nM for chloroquine; 4.8, 2.6, 9.0, 6.9 and 5.8 nM for mefloquine; 21.3, 8.3, 41.9, 49.6 and 40.1 nM for quinine, 129.9, 47.3, 109.3, 30.6 and 95.4 nM for cycloguanil, and 45.4, 47.4, 40.2, 66.3 and 36.0 nM for pyrimethamine. IC50 (nM) for Gombak A, Gombak C, ST 9, ST 12, ST 85 and ST 148 isolates after 12 months of continuous in vitro culture were, respectively: 477.0, 492.3, 367.1, 809.4, 566.5 and 341.8 for chloroquine; 2.9, 11.1, 8.5, 16.9, 5.3 and 4.2 for mefloquine; 6.2, 58.3, 52.7, 36.7, 31.8 and 26.2 for quinine; 154.5, 57.2, 130.3, 94.2, 81.4 and 102.9 for cycloguanil, 26.9, 24.9, 43.8, 31.0, 14.1 and 56.7 for pyrimethamine. Before the 12-month culture they were 472.3, 452.9, 352.7, 773.7, 702.7 and 322.7 nM for chloroquine; 2.6, 13.2, 8.5, 17.2, 5.0 and 4.0 nM for mefloquine; 6.2, 85.4, 53.9, 38.5, 35.8 and 38.5 nM for quinine; 106.8, 74.3, 112.4, 89.8, 91.8 and 103.3 nM for cycloguanil, and 26.9, 31.4, 47.0, 28.1, 14.9 and 56.7 nM for pyrimethamine. Thus none of these isolates differed in their original susceptibilities after either of these procedures.
    Matched MeSH terms: Cryopreservation*
  8. Philip N, Garba B, Neela VK
    Appl Microbiol Biotechnol, 2018 Jul;102(13):5427-5435.
    PMID: 29736823 DOI: 10.1007/s00253-018-9047-9
    Preservation of leptospiral cultures is tantamount to success in leptospiral diagnostics, research, and development of preventive strategies. Each Leptospira isolate has imperative value not only in disease diagnosis but also in epidemiology, virulence, pathogenesis, and drug development studies. As the number of circulating leptospires is continuously increasing and congruent with the importance to retain their original characteristics and properties, an efficient long-term preservation is critically needed to be well-established. However, the preservation of Leptospira is currently characterized by difficulties and conflicting results mainly due to the biological nature of this organism. Hence, this review seeks to describe the efforts in developing efficient preservation methods, to discover the challenges in preserving this organism and to identify the factors that can contribute to an effective long-term preservation of Leptospira. Through the enlightenment of the previous studies, a potentially effective method has been suggested. The article also attempts to evaluate novel strategies used in other industrial and biotechnological preservation efforts and consider their potential application to the conservation of Leptospira spp.
    Matched MeSH terms: Cryopreservation/methods*
  9. 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*
  10. Lee SH, Looi CY, Chong PP, Foo JB, Looi QH, Ng CX, et al.
    Curr Stem Cell Res Ther, 2021;16(5):551-562.
    PMID: 32988356 DOI: 10.2174/1574888X15666200928110923
    Mesenchymal Stem Cells (MSCs) are adult stem cells that are gaining worldwide attention for their multi-potential use in tissue engineering-based regenerative medicine. They can be obtained from numerous sources and one of the excellent sources is the dental tissue, such as Stem cells that are extracted from the Human Exfoliated Deciduous teeth (SHED). SHED are considered ideal due to their inherent characteristics, including the capability to proliferate quickly with minimal oncogenesis risk, multipotency capacity and their ability to suppress the immune system. On top of these positive cell traits, SHED are easily accessible with the patient's safety assured, posing less ethical issues and could also provide a sufficient number of cells for prospective clinical uses. This is primarily attributed to their ability to differentiate into multiple cell linages, including osteoblasts, odontoblasts, neuronal cells, adipocytes, as well as endothelial cells. Albeit SHED having a bright future, there still remains an obstacle to develop reliable experimental techniques to retain the long-term regeneration potential of the stem cells for prospective research and clinical applications. Therefore, this review aims to describe the various isolation, expansion and cryopreservation techniques used by researchers in this stem cell field. Optimization of these techniques is crucial to obtain distinct SHED culture with preserved stem cell properties, which enable more reproducible results that will be the key for further stem cell therapy development.
    Matched MeSH terms: Cryopreservation*
  11. Suppiah J, Nadaraju S, Hamzah S, Chee HY
    Trop Biomed, 2020 Jun 01;37(2):282-287.
    PMID: 33612798
    Storage of dengue virus (DENV) culture stocks in -80°C is a common laboratory practice to maintain the viability of the virus for long-term usage. However, the efficiency of this method could still be hindered by multiple factors. In our laboratory, we observed a constant and substantial deterioration in the titer of DENV in Vero culture supernatant stored in -80°C. Such incident had badly hampered the laboratory work and prompted an investigation to determine the cause. DENV isolates representing all four serotypes were propagated and the culture supernatants were harvested and stored in aliquots of original stock and 10 fold dilutions (10-1 -10-4). DENV titer in these stocks was determined prior to storage and reassessed on the third and sixth month of storage by focus forming unit assay (FFUA). The result demonstrated a constant preservation of titer ranging from 104 ffu/ml to 105 ffu/ml in the diluted DENV virus culture stocks of 10-1, and 10-2 of DENV1-4, a minor reduction of titer from 103 ffu/ml to 102 ffu/ml at dilution 10-3 for DENV4 only and complete deterioration in undiluted culture stock and lower dilution (10-4) within 6 months of storage in -80°C for all serotypes. It is recommended that propagated DENV in Vero cells are stored in 10 fold dilutions as compared to the original form to preserve the titer for long-term usage.
    Matched MeSH terms: Cryopreservation*
  12. Tan YC, Mustangin M, Rosli N, Wan Ahmad Kammal WSE, Md Isa N, Low TY, et al.
    Cryobiology, 2024 Mar;114:104843.
    PMID: 38158171 DOI: 10.1016/j.cryobiol.2023.104843
    Coolant-assisted liquid nitrogen (LN) flash freezing of frozen tissues has been widely adopted to preserve tissue morphology for histopathological annotations in mass spectrometry-based spatial proteomics techniques. However, existing coolants pose health risks upon inhalation and are expensive. To overcome this challenge, we present our pilot study by introducing the EtOH-LN workflow, which demonstrates the feasibility of using 95 % ethanol as a safer and easily accessible alternative to existing coolants for LN-based cryoembedding of frozen tissues. Our study reveals that both the EtOH-LN and LN-only cryoembedding workflows exhibit significantly reduced freezing artifacts compared to cryoembedding in cryostat (p 
    Matched MeSH terms: Cryopreservation/methods
  13. Ali J, AlHarbi NH, Ali N
    Methods Mol Biol, 2017;1568:3-20.
    PMID: 28421485 DOI: 10.1007/978-1-4939-6828-2_1
    This chapter describes the development of the science of cryopreservation of gametes and embryos of various species including human. It attempts to record in brief the main contributions of workers in their attempts to cryopreserve gametes and embryos. The initial difficulties faced and subsequent developments and triumphs leading to present-day state of the art are given in a concise manner. The main players and their contributions are mentioned and the authors' aim is to do justice to them. This work also attempts to ensure that credit is correctly attributed for significant advances in gamete and embryo cryopreservation. In general this chapter has tried to describe the historical development of the science of cryopreservation of gametes and embryos as accurately as possible without bias or partiality.
    Matched MeSH terms: Cryopreservation
  14. Boey KPY, Zhu P, Tan H, Abdullah MAB, Tang KF, Li MM, et al.
    Transfus Med, 2022 Feb;32(1):82-87.
    PMID: 34862686 DOI: 10.1111/tme.12834
    OBJECTIVE: To evaluate the effects of cryopreservation in post-thaw umbilical cord blood units for the survivability of Gram-positive bacteria strains.

    BACKGROUND: Microbial screening is required for all cord blood units (CBUs). Four gram-positive contaminants were documented to survive cryopreservation poorly and isolation of other contaminants were reported.

    METHODS: Forty-eight contaminated CBUs detected with either Staphylococcus epidermidis, Corynebacterium species, Peptostreptococcus or Streptococcus species before cryopreservation were used in this study. CBUs were processed, DMSO-infused and microbial screened before cryopreservation. Post-thaw microbial screening was achieved using 1 and 10 ml inoculants in BACTEC culture bottles. Positive bottles were subjected for microbial identification and results were compared with those from pre-freeze.

    RESULTS: A higher rate of microbial contamination was found using the 10 ml inoculant. Screening of 11 CBUs did not detect any contaminants while 30 CBUs screened detected more than one unknown contaminants and majority of contaminants were identified to be gram-negative species.

    CONCLUSION: A higher inoculation volume used at post-thaw for microbial screening improves contamination detection but leads to the loss of precious cord blood. Some contaminants did not survive cryopreservation or were not identified due to their low microbial levels. Contrasting contaminants found at post-thaw suggest the improvements made in detection and identification of contaminants over the years.

    Matched MeSH terms: Cryopreservation
  15. Gantait S, Sinniah UR, Suranthran P, Palanyandy SR, Subramaniam S
    Protoplasma, 2015 Jan;252(1):89-101.
    PMID: 24893588 DOI: 10.1007/s00709-014-0660-x
    In the present study, polyembryoids of oil palm (Elaeis guineensis Jacq.) were cryopreserved with successful revival of 68 % for the first time using the droplet vitrification technique. Excised polyembryoids (3-5-mm diameter) from 3-month-old in vitro cultures were pre-cultured for 12 h in liquid Murashige and Skoog medium supplemented with 0.5 M sucrose. The polyembryoids were osmoprotected in loading solution [10% (w/v) dimethyl sulphoxide (DMSO) plus 0.7 M sucrose] for 30 min at room temperature and then placed on aluminium strips where they were individually drenched in chilled droplets of vitrification solution (PVS2) [30% (w/v) glycerol plus 15% (w/v) ethylene glycol (EG) plus 15% (w/v) DMSO plus 0.4 M sucrose] for 10 min. The aluminium strips were enclosed in cryovials which were then plunged quickly into liquid nitrogen and kept there for 1 h. The polyembryoids were then thawed and unloaded (using 1.2 M sucrose solution) with subsequent transfer to regeneration medium and stored in zero irradiance. Following for 10 days of storage, polyembryoids were cultured under 16 h photoperiod of 50 μmol m(-2) s(-1) photosynthetic photon flux density, at 23 ± 1 °C. Post-thaw growth recovery of 68% was recorded within 2 weeks of culture, and new shoot development was observed at 4 weeks of growth. Scanning electron microscopy revealed that successful regeneration of cryopreserved polyembryoids was related to maintenance of cellular integrity, presumably through PVS2 exposure for 10 min. The present study demonstrated that cryopreservation by droplet vitrification enhanced the regeneration percentages of oil palm in comparison with the conventional vitrification method previously reported.
    Matched MeSH terms: Cryopreservation/methods*
  16. 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*
  17. 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*
  18. Nang CF, Osman K, Budin SB, Ismail MI, Jaffar FH, Mohamad SF, et al.
    Andrologia, 2012 May;44 Suppl 1:447-53.
    PMID: 21806660 DOI: 10.1111/j.1439-0272.2011.01203.x
    Liquid nitrogen preservation in remote farms is a limitation. The goal of this study was to determine optimum temperature above freezing point for bovine spermatozoa preservation using bovine serum albumin (BSA) as a supplementation. Pooled semen sample from three ejaculates was subjected to various BSA concentration (1, 4, 8 and 12 mg ml(-1)), before incubation in different above freezing point temperatures (4, 25 and 37 °C). Viability assessment was carried out against time from day 0 (fresh sample) until all spermatozoa become nonviable. Optimal condition for bovine spermatozoa storage was at 4 °C with 1 mg ml(-1) BSA for almost 7 days. BSA improved bovine spermatozoa viability declining rate to 44.28% at day 4 and 57.59% at day 7 compared to control, with 80.54% and 98.57% at day 4 and 7 respectively. Increase in BSA concentration did not improve sperm viability. Our results also confirmed that there was a strong negative correlation between media osmolarity and bovine spermatozoa survival rate with r = 0.885, P < 0.0001. Bovine serum albumin helps to improve survival rate of bovine spermatozoa stored above freezing point.
    Matched MeSH terms: Cryopreservation*
  19. Chua SP, Normah MN
    Cryo Letters, 2011 Nov-Dec;32(6):506-15.
    PMID: 22227711
    This paper reports the cryopreservation of Nephelium ramboutan-ake shoot tips derived from in vitro shoot multiplication and in vitro seed germination using vitrification. Preculture with either 0.5 M sucrose for 2 days or a combination of 0.3 M sucrose and 0.5 M glycerol for 3 days enhanced dehydration tolerance and resulted in the highest survival of shoot tips; however, none of the shoot tips withstood liquid nitrogen (LN) exposure. The use of a lower temperature (0 degree C) during exposure to plant vitrification solution (PVS2) led to higher survival of shoot tips, compared to exposure at 25 degree C. The survival percentage of shoot tips exposed to PVS2 for up to 20 min at 0°C was 83.3 percent. It was only 53.3 percent when shoot tips were exposed to PVS2 at 25 degree C for 5 min. The importance of vitamin C for reducing oxidative stress in shoots tips was demonstrated. The addition of 0.28 mM vitamin C during critical steps of the vitrification process resulted in a high survival (96.7 percent) without LN exposure, compared to 73.3 percent for shoot tips not treated with vitamin C. Moreover, 3.3 percent shoot tips withstood LN exposure when vitamin C was added during the loading step. This result suggests that cryopreservation is possible for this tropical, recalcitrant seeded tree species.
    Matched MeSH terms: Cryopreservation*
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