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iPSC-hDPSC
Tissue
第三臼齒
Disease
一般捐贈者

Documents

iPSC-hDPSC

Cell line · Product number 300622

The iPSC-hDPSC cell line represents a cutting-edge model derived from human dental pulp stem cells (hDPSCs). These cells originate from dental pulp tissue...

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01General information

Description

iPSC-hDPSC 細胞系是一種源自人類牙髓幹細胞(hDPSCs)的前沿模型。這些細胞源自牙髓組織,並經過酶消化處理以分離出貼壁細胞群。 當細胞匯合度達到 70-80% 時,便進行傳代培養,並於第 1 代(Passage 1)進行冷凍保存。此基礎步驟對於維持細胞完整性,以及確保幹細胞在未來應用中的存活能力至關重要。

將 hDPSC 重編程為誘導性多能幹細胞(iPSC)是一項重大進展,是透過環狀DNA重編程技術實現的。 此方法採用載有山中因子(Oct-4、Sox-2 和 Klf-4)的載體,並結合 p53 反義序列、EBNA-1 以及紅色螢光蛋白標記。 外源體方法的特別優勢在於,它避免了重編程因子整合到基因組中,從而維持了 iPSC 的遺傳穩定性。由此產生的 iPSC-hDPSC 細胞系展現出多能性特徵,使其成為再生醫學、疾病模型建立及藥物發現研究中極具價值的工具。

Tissue
第三臼齒
Disease
一般捐贈者
Applications
疾病建模、藥物研發與毒性測試、再生醫學、基因研究、發育生物學研究

02Classification

Organism
Human
Growth properties
Adherent
GMO status
GMO-S1
GMO status details
GMO-S1: This human iPSC line (iPSC-hDPSC) contains episomal plasmids encoding OCT4, SOX2, KLF4 and c-MYC, supporting reprogramming of dental stromal cells. The constructs are maintained without viral sequences. This classification applies only within Germany and may differ elsewhere.
Biosafety level
1

03Characteristics

Morphology
IPSC,邊界明確的聚落
Cell type
幹細胞

04Identifiers

Citation
iPSC-hDPSC(Cytion 型號 300622)
NCBI TaxID
9606
Cellosaurus
CVCL_B7DW

05Handling and culture

Culture medium
mTESR
Incubation atmosphere

37°C,5% CO₂,加濕環境。

Dissociation reagent
ReLeSR
Subculturing

為改善細胞脫附與重新接種的流程,請依照以下步驟操作:

  1. 移除用過的培養基,並用不含鈣和鎂的 PBS 沖洗細胞。
  2. 加入脫附溶液,並依照製造商的指示靜置作用。
  3. 小心地向細胞中加入新鮮培養基。
  4. 輕柔地將細胞解離,以確保損傷降至最低。
  5. 在全新的 6 孔培養板表面塗佈維特羅內克汀,並將解離後的細胞接種至其中兩個孔中,以進行後續培養。
Seeding density

為確保細胞生長狀況最佳且實驗結果具一致性,建議在細胞適應培養條件後,將接種密度設定為 5,000 至 10,000 個細胞/cm²。

Fluid renewal
每日
Post-thaw recovery
快速
Thawing and culturing
  1. Confirm that the vial remains deeply frozen upon delivery, as cells are shipped on dry ice to maintain optimal temperatures during transit.

  2. Upon receipt, either store the cryovial immediately at temperatures below -150°C to ensure the preservation of cellular integrity, or proceed to step 3 if immediate culturing is required.

  3. For immediate culturing, swiftly thaw the vial by immersing it in a 37°C water bath with clean water and an antimicrobial agent, agitating gently for 40-60 seconds until a small ice clump remains.

  4. Perform all subsequent steps under sterile conditions in a flow hood, disinfecting the cryovial with 70% ethanol before opening.

  5. Carefully open the disinfected vial and transfer the cell suspension into a 15 ml centrifuge tube containing 8 ml of room-temperature culture medium, mixing gently.

  6. Centrifuge the mixture at 300 x g for 3 minutes to separate the cells and carefully discard the supernatant containing residual freezing medium.

  7. Gently resuspend the cell pellet in 10 ml of fresh culture medium. For adherent cells, divide the suspension between two T25 culture flasks; for suspension cultures, transfer all the medium into one T25 flask to promote effective cell interaction and growth.

  8. Adhere to established subculture protocols for continued growth and maintenance of the cell line, ensuring reliable experimental outcomes.

06Quality control and cryopreservation

Sterility

透過基於 PCR 的檢測及基於發光原理的支原體檢測方法,排除支原體污染的可能性。

為確保無細菌、真菌或酵母菌污染,每日均會對細胞培養物進行目視檢查。

Freeze medium
作為冷凍保存培養基,我們使用 80% 胎牛血清(FBS)+ 10% 基礎培養基 + 10% 二甲基亞砜(DMSO)來維持細胞存活率;或使用 CM-1(Cytion 型號 800100)以獲得更佳的冷凍保護效果,在維持多能性與細胞完整性的同時,防止不必要的分化。
Storage conditions

若需長期保存,請將試管置於溫度約為 −150 至 −196 °C 的氣相液氮中。僅在轉移至液氮之前,可將試管暫時存放於 −80 °C 環境中,作為短期過渡步驟。

Shipping conditions

冷凍保存的細胞系會以乾冰冷藏,並裝於經驗證的隔熱包裝中,包裝內含有足夠的冷媒,以確保在運輸過程中溫度維持在約 −78 °C。收到貨物後,請立即檢查容器,並盡快將試管轉移至適當的儲存場所。

07Donor

Ethnicity
高加索人