Effect of different cryopreservation media on human nucleus pulposus cells' viability and trilineage potential.
Effect of different cryopreservation media on human nucleus pulposus cells' viability and trilineage potential.
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DOI：
10.1002/jsp2.1140
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发表时间：
2021-03
期刊：
影响因子：
3.7
通讯作者：
Gantenbein B
中科院分区：
文献类型：
--
作者：
Croft AS;Guerrero J;Oswald KAC;Häckel S;Albers CE;Gantenbein B
简介：在许多国家，下腰痛(LBP)是导致残疾的一个重要原因，全世界有超过5亿人受到影响。过去，在人类椎间盘(IVD)的髓核(NP)内发现了祖细胞。然而，在细胞治疗的背景下，关于冷冻保存和扩增对这里所称的“异源”人NP细胞的影响，以及商业上可用的冷冻保存介质是否在细胞活力方面比“常用”介质更有效，人们知之甚少。材料：本研究收集了4例创伤患者(年龄40.5 ± 14.3 )和2例退行性脑室内畸形患者(年龄分别为2 4和46 )脊柱外科手术患者的hNCs。为了分离hNPC，用温和的两步法消化组织。扩增后，分离传代2~5代的人神经干细胞，在−15 0℃下冷冻保存1 周，或分化为成骨、成脂或成软骨细胞系21天。用5种不同的培养液进行冷冻保存，比较它们对细胞活力和分化潜能的影响。流式细胞仪检测细胞存活率，定量聚合酶链式反应和组织学分析检测细胞分化潜能。结果：人鼻咽癌细胞冷冻保存1 周后，细胞活力在所有条件下均可比，即与所用的冷冻保存液(82.3 ± 为细胞活力的0.8%)无关。此外，创伤患者的hNPC显示了一些成脂和成软骨分化的证据，在较低水平，这一点和成骨分化的证据可以用来自退变的椎间盘的hNPC来证实。此外，在大多数被测试的病例中，冷冻保存并不影响细胞的分化潜力。结论：在细胞活力和hNPC三联体分化潜能的总体维持方面，“常用的”冷冻保存液似乎与商用的冻存液一样好。分离、扩增人髓核细胞群，然后用5种不同的冻存介质中的一种在−150°C中冷冻保存，并/或评估细胞的三倍体分化潜能。细胞的活性在测试的冷冻保存液中没有差异，hNPC显示有成骨、成脂和成软骨分化的证据。
Introduction: Low back pain (LBP) is a significant cause of disability in many countries, affecting more than half a billion people worldwide. In the past, progenitor cells have been found within the nucleus pulposus (NP) of the human intervertebral disc (IVD). However, in the context of cell therapy, little is known about the effect of cryopreservation and expansion on here called “heterogenic” human NP cells (hNPCs), and whether commercially available cryopreservation media are more efficient than “commonly used” media in terms of cell viability. Materials: In this study, hNPCs from four trauma patients (age 40.5 ± 14.3 years) and two patients with degenerated IVDs (age 24 and 46 years), undergoing spinal surgery, were collected. To isolate hNPCs, the tissue was digested with a mild two‐step protocol. After subsequent expansion, hNPCs at passages 2‐5 were separated and either cryo‐preserved for 1 week at −150°C or differentiated into osteogenic, adipogenic, or chondrogenic lineages for 21 days. Cryopreservation was performed with five different media to compare their effect on the cell's viability and differentiation potential. Cell viability was determined with flow cytometry using propidium iodide and the trilineage differentiation potential was assessed by quantitative polymerase chain reaction and histological analysis. Results: After 1 week of cryopreservation, the hNPC's cell viability was comparable for all conditions, that is, independent of the cryopreservation medium used (82.3 ± 0.8% of cell viability). Furthermore, hNPCs from trauma patients showed some evidence for adipogenic and chondrogenic differentiation and at lower levels, this and evidence of osteogenic differentiation could be confirmed with hNPCs from degenerated discs. Moreover, cryopreservation did not affect the cell's differentiation potential in the majority of the cases tested. Conclusion: “Commonly used” cryopreservation media seem to perform just as well as commercially available media in terms of cell viability and the overall maintenance of the hNPCs trilineage differentiation potential. Human nucleus pulposus cell populations (hNPCs) were isolated, expanded, and then cryo‐preserved at −150°C with one of five different cryopreservation media and/or the cells' trilineage differentiation potential was assessed. The cells' viability did not differ between the cryopreservation media tested and hNPCs showed evidence for osteogenic, adipogenic and chondrogenic differentiation.
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影响因子：
2.8
作者：
Haschtmann, Daniel;Ferguson, Stephen J.;Stoyanov, Jivko V.
通讯作者：
Stoyanov, Jivko V.
DOI：
10.1007/978-1-4939-6921-0_5
发表时间：
2017-01-01
期刊：
STEM CELL BANKING: CONCEPTS AND PROTOCOLS
影响因子：
--
作者：
Hunt, Charles J.
通讯作者：
Hunt, Charles J.
DOI：
10.1097/01.blo.0000198726.62514.75
发表时间：
2006-02-01
影响因子：
4.2
作者：
Deyo, Richard A.;Mirza, Sohail K.
通讯作者：
Mirza, Sohail K.
影响因子：
2.9
作者：
CASSIDY, JJ;HILTNER, A;BAER, E
通讯作者：
BAER, E
影响因子：
2.8
作者：
Brown, Sharon J.;Turner, Sarah A.;Roberts, Sally
通讯作者：
Roberts, Sally