Human umbilical cord mesenchymal cells from 4 donors cultured from P1 or P2 to replicative senescence. Human umbilical cord mesenchymal cells from 4 donors cultured from P1 or P2 to replicative senescence

To characterize the growth kinetics and transcriptome drift of human umbilical cord perivascular cells (HUCPVCs), cells were recovered from cryopreservation and cultured in chemically defined xeno- and serum-free media until replicative senescence. Growth kinetics and transcriptome profiles were analyzed at each passage. Overall design: Aliquots of 1 million cells from 4 donor populations of HUCPVCs, provided by Tissue Regeneration Therapeutics (TRT), Inc. (Toronto, Canada), were thawed from cryopreservation and subject to independent, continuous in vitro culture in serum- and xeno-free media until they reached replicative senescence. Senescence was defined as sub-confluence 4 or more weeks after seeding in the current culture vessel. Cells were lifted at 70-80% confluence in culture vessels, denoting the end of each passage, and re-seeded. RNA was extracted from the remaining cell collection and analyzed by Affymetrix Human Genome U133A 2.0 Array. Information on batches of experimental culture and GeneChip processing time frames, encompassing variability of laboratory conditions and reagent lots, is included.

为表征人脐带周血管细胞（human umbilical cord perivascular cells, HUCPVCs）的生长动力学与转录组漂移特征，本研究将细胞从冷冻保存中复苏，接种于化学成分限定、无异种源且无血清的培养基中连续培养，直至细胞进入复制性衰老（replicative senescence）。研究过程中，于每一代传代节点对细胞的生长动力学与转录组图谱进行分析。整体实验设计：由加拿大多伦多的组织再生治疗公司（Tissue Regeneration Therapeutics, TRT, Inc.）提供的4个HUCPVC供体细胞群，各取100万细胞分装后从冷冻保存中复苏，分别开展独立的连续体外培养，培养体系均采用无血清、无异种源的化学成分限定培养基，直至细胞达到复制性衰老。本研究中将复制性衰老定义为：在当前培养容器中接种后，细胞汇合度未达饱和（即未铺满培养容器底面积）且培养时长达到4周及以上。当细胞在培养容器中生长至70%-80%汇合度时，对细胞进行收获并传代，此为每一代培养的结束节点，随后将细胞重新接种至新的培养容器。收集剩余细胞提取总RNA，通过Affymetrix人类基因组U133A 2.0芯片（Affymetrix Human Genome U133A 2.0 Array）进行转录组分析。本研究同时收录了实验培养批次与GeneChip芯片处理时间窗口的相关信息，以覆盖实验室操作条件、试剂批次等带来的变量差异。