Cultivation and cryopreservation of cord tissue MSCs with Cord Blood AB Plasma

Manasi Taiwadekarr, D. Kadekar, Sonal Rangole, Nikhat Khan, V. Kale, L. Limaye
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引用次数: 6

Abstract

Neonatal tissues, cord and placenta, are explored as alternate sources of mesenchymal stem cells (MSCs) for their therapeutic applications. Conventionally, MSCs isolated from cord tissues are maintained and propagated in FBS containing medium for promotion of growth and survival of cells. However, for therapeutic use, FBS use is not encouraged as it is of animal origin. Thus, there is a need for replacement of FBS by equally potent and clinically acceptable cost effective sources. The current study is designed to compare the effect of cord blood plasma (CBP) with MSC qualified FBS (M FBS) during culture and cryopreservation of MSCs. MSCs were isolated from cord and placenta and propagated in either M FBS or CBP. The efficiency of the cultures was analyzed by growth curve, morphology, phenotype and functionality. The cryo-protective role of the CBP was evaluated by using it in freezing medium of MSCs. Our data showed that CBP is equivalent to M FBS for culturing placental MSCs with respect to the phenotype, proliferation rate and differentiation to various lineages. However, cord MSCs displayed slow growth rate and reduction in surface expression of CD105 marker in CBP, whereas, the other parameters were comparable. Freezing of MSCs with CBP resulted in reduction of the late apoptotic and necrotic population. Thus, CBP imparts superior protection against cryogenic insults, and appears to be a valuable substitute to M FBS for cultivation and freezing of MSCs.
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脐带血AB血浆培养及超低温保存脐带血MSCs
新生儿组织,脐带和胎盘,被探索作为间充质干细胞(MSCs)的治疗应用的替代来源。通常,从脐带组织中分离的间充质干细胞在含有FBS的培养基中维持和繁殖,以促进细胞的生长和存活。然而,对于治疗用途,不鼓励使用FBS,因为它是动物来源的。因此,有必要用同样有效和临床可接受的成本效益来源替代FBS。本研究旨在比较脐带血血浆(CBP)与MSC合格FBS (M FBS)在MSCs培养和冷冻保存过程中的作用。从脐带和胎盘中分离MSCs,在M - FBS或CBP中繁殖。通过生长曲线、形态、表型和功能分析培养效率。在骨髓间充质干细胞冷冻培养基中观察CBP的低温保护作用。我们的数据显示,CBP在表型、增殖率和向不同谱系的分化方面与M FBS在培养胎盘间充质干细胞方面相当。然而,脐带间充质干细胞在CBP中表现出生长速度缓慢和CD105标记物表面表达减少,而其他参数具有可比性。用CBP冷冻MSCs可减少晚期凋亡和坏死细胞数量。因此,CBP对低温损伤具有优越的保护作用,似乎是mfbs培养和冷冻MSCs的有价值的替代品。
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